@article{SebaldBuecherOlbrichetal.1968,
  author    = {Sebald, Walter and B{\"u}cher, T. and Olbrich, B. and Kaudewitz, F.},
  title     = {Electrophoretic pattern of and amino acid incorporation in vitro into the insoluble mitochondrial protein of neurospora crassa wild type and mi-1 mutant},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62926},
  year      = {1968},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{NeupertSebaldSchwabetal.1969,
  author    = {Neupert, W. and Sebald, Walter and Schwab, A. J. and Pfaller, A. and B{\"u}cher, T.},
  title     = {Puromycin sensitivity of ribosomal label after incorporation of \(^{14}\)C-labelled amino acids into isolated mitochondria from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62899},
  year      = {1969},
  abstract  = {Radioactive amino acids were incorporated into isolated mitochondria from Neurospora crassa. Then the mitochondrial ribosomes were isolated and submitted to density gradient centrifugation. A preferential labelling of polysomes was observed. However, when the mitochondrial suspension was treated with puromycin after amino acid incorporation, no radioactivity could be detected in either the monosomes or the polysomes. The conclusion is drawn that isolated mitochondria under these conditions do not incorporate significant amounts of amino acids into proteins of their ribosomes.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldSchwabBuecher1969,
  author    = {Sebald, Walter and Schwab, A. J. and B{\"u}cher, T.},
  title     = {Cycloheximide resistant amino acid incorporation into mitochondrial protein from Neurospora crassa in vivo},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62900},
  year      = {1969},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldHofstoetterHackeretal.1969,
  author    = {Sebald, Walter and Hofst{\"o}tter, T. and Hacker, D. and B{\"u}cher, T.},
  title     = {Incorporation of amino acids into mitochondrial protein of the flight muscle of Locusta migratoria in vitro and in vivo in the presence of cycloheximide},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62919},
  year      = {1969},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{NeupertSebaldSchwabetal.1969,
  author    = {Neupert, W. and Sebald, Walter and Schwab, A. J. and Massinger, P. and B{\"u}cher, T.},
  title     = {Incorporation in vivo of \(^{14}\)C-labelled amino acids into the proteins of mitochondrial ribosomes from Neurospora crassa sensitive to cycloheximide and insensitive to Chloramphenicol},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62884},
  year      = {1969},
  abstract  = {Radioactive amino acids were incorporated in vivo into N eurospora crassa cells, and the mitochondrial ribosomes were isolated. The incorporation of radioactivity into the proteins of these ribosomes was inhibited by cycloheximide, but not by chloramphenicol. It is therefore concluded that these proteins are synthesized on the cycloheximide sensitive and chloramphenicol insensitive cytoplasmic ribosomes.},
  subject      = {Biochemie},
  language  = {en}
}
@article{BirkmayerSebaldBuecher1969,
  author    = {Birkmayer, G. D. and Sebald, Walter and B{\"u}cher, Th.},
  title     = {Cytochrom-Oxydase und ein an diese assoziiertes, markiertes Protein aus 14C-markierten Mitochondrien von Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82135},
  year      = {1969},
  abstract  = {no abstract available},
  subject      = {Physiologische Chemie},
  language  = {de}
}
@article{SebaldNeupertBirkmayer1970,
  author    = {Sebald, Walter and Neupert, W. and Birkmayer, G. D.},
  title     = {Internal and external contributions to the biogenesis of mitochondrial proteins},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62876},
  year      = {1970},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{WeissSebaldBuecher1971,
  author    = {Weiss, H. and Sebald, Walter and B{\"u}cher, T.},
  title     = {Cycloheximide resistant incorporation of amino acids into a polypeptide of the cytochrome oxidase of Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62866},
  year      = {1971},
  abstract  = {Radioaetive leueine was ineorporated by N eurospora crassa mitoehondria in vivo in the presence of cyeloheximide. When the membrane protein of these mitochondria was ehromatographieally separated on oleyl polymethaerylie aeid resin, \& nurober of fraetions were obtained whieh differ with respeet to their eontents of radioaetivity and eytoehromes. The highest speeifie radioaetivity was found in the fraction eontaining eytoehrome aa3• This fraetion proved to be a pure and enzymatically aetive cytoehrome oxidase. Its ratio of absorbanee at 280 nm (ox)/ 443 nm (red.) was 2.1. By means of sodium dodeeylsulfate gel-electrophoresis, this enzymewas separated into five polypeptides with molecular weights of 30000, 20000, 13000, 10000, and 8000. Only the polypeptide with the molecular weight 20000 displayed a high specific radioaetivity.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SchwabSebaldWeiss1972,
  author    = {Schwab, A. J. and Sebald, Walter and Weiss, H.},
  title     = {Different pool sizes of the precursor polypeptides of cytochrome oxidase from Neurospora crassa.},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62841},
  year      = {1972},
  abstract  = {Pulse-labelling experiments with growing Neurospora crassa revealed that the polypeptides composing the protein moiety of a cytochrome oxidase preparation are derived from at least four independent pools of precursor polypeptides. The pool sizes range from 2 ° f 0 to 25 °/0 of the amount of the corresponding polypeptide present in cytochrome oxidase. The smallest pool is assigned to a polypeptide of mitochondrial origm. Serial pools were found for one of the polypeptides.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldWeissJackl1972,
  author    = {Sebald, Walter and Weiss, H. and Jackl, G.},
  title     = {Inhibition of the assembly of cytochrome oxidase in Neurospora crassa by chloramphenicol},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62852},
  year      = {1972},
  abstract  = {Cytochrome oxidasewas prepared from Neurospora crassa by chromatography on oleyl polymethacrylic acid resin and separated into seven polypeptides by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate. Incorporation oflabelled amino acids into the single polypeptideswas investigated after a pulse labelling in the absence and presence of chloramphenicol, and afterwashing out the inhibitor. Chloramphenicol (4 mg/ml) inhibited amino acid incorporation into all polypeptides 90-95\%• while labeHing of the whole membrane protein was inhibited only 30\%• Mter washing out the inhibitor and further growth of the cells. the four smaller polypeptides were highly labelled, whereas the other polypeptides showed only a. small increase in radioactivity. It is concluded that the four small-sized polypeptides of cytochrome oxidase are synthesized but not integrated into the functional enzyme under the action of chloramphenicol.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SchwabSebaldWeiss1972,
  author    = {Schwab, A. J. and Sebald, Walter and Weiss, H.},
  title     = {Schnelle Markierung eines mitochondrial synthetisiertem Polypeptids einer Cytochromoxidasen-Pr{\"a}paration aus Neurospora},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-84206},
  year      = {1972},
  abstract  = {no abstracts available},
  subject      = {Physiologische Chemie},
  language  = {de}
}
@article{SebaldWeissJackl1972,
  author    = {Sebald, Walter and Weiss, H. and Jackl, G.},
  title     = {{\"U}ber die Abh{\"a}ngigkeit des Zusammenbaus der Cytochromoxidase von der Anwesenheit der Produkte der mitochondrialen Proteinsynthese},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-84192},
  year      = {1972},
  abstract  = {no abstract available},
  subject      = {Physiologische Chemie},
  language  = {de}
}
@article{SebaldMachleidtOtto1973,
  author    = {Sebald, Walter and Machleidt, W. and Otto, J.},
  title     = {Products of mitochondrial protein synthesis in Neurospora crassa. Determination of equimolar amounts of three products in cytochrome oxidase on the basis of amino-acid analysis},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62827},
  year      = {1973},
  abstract  = {Cytochrome oxidase isolated from N eurospora crassa was resolved into seven protein eomponents by eleetrophoresis in polyaerylamide gels eontaining sodium dodeeylsulfate. The apparent molecular weights were determined tobe 41000, 28500, 21000, 16000, 14000, 11500 and 10000 for the eomponents 1, 2, 3, 4, 5, 6, and 7, respectively. The components 1, 2 and 3 are synthesized on mitochondrial ribosomes as shown by the incorporation of radioactive amino aeids in the presenee of cyeloheximide. Amino-acidanalysis of the isolated components 1, 2 and 3 revealed a high content of apolar amino acids and a low eontent of basic amino aeids compared to an average amino-aeid eomposition of components 4-7. Components 1, 2 and 3 eontribute 27.9°/0, 18°/0 and 14.2°/0 to the whole eytoehrome oxidase protein. This was calculated from the contributions of the single eomponents to the totalleueine eontent of the enzyme and the leueine eontents (nmol leueine per mg protein) of the single eomponents as determined by amino-aeid analysis. Equimolar relations of the components 1, 2 and 3 are found by dividing the amounts of protein by their apparent molecular weights. A stoichiometry of 1:1:1 results assuming a minimal molecular weight of 150000 for the whole cytochrome oxidase protein. On the basis of the heme a content a molecular weight of about 70000 per heme group was determined, using an absorption coeffieient L1e605 (redueed minus oxidized) of 12 mM-1 cm-1• It is concluded that the smallest structural unit of eytochrome oxidase contains two heme groups.},
  subject      = {Biochemie},
  language  = {en}
}
@article{WeissSebaldSchwabetal.1973,
  author    = {Weiss, H. and Sebald, Walter and Schwab, A. J. and Kleinow, W. and Lorenz, B.},
  title     = {Contribution of mitochondrial and cytoplasmic protein synthesis to the formation of cytochrome b and cytochrome aa\(_3\)},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62835},
  year      = {1973},
  abstract  = {A cytochrome b preparation from Neurospora crassa mitochondria is found to consist of three polypeptides (apparent molecular weight 10 000, 11 000 and 32 000), a cytochrome aa3 preparation of six to seven polypeptides (apparent molecular weight 8 000, 11 000, 13 000, 18 000, 28 000 and 36 000). Selective incorporation of radioactive amino acids by eilher mitochondrial protein synthesis when the cytoplasmic one is blocked or by the cytoplasmic protein synthesis, when the mitochondrial one is blocked, indicates that one cytochrome b polypeptide (mw 32 000) and one to three cytochrome aa3 polypeptides (mw 36 000, 28 000 and 18 000) are mitochondrial translation products, the other cytochrome b and cytochrome aa3 polypeptides cytoplasmic translation products. The delayed appearance of labeling in the cytochrome b and cytochrome aa3 polypeptides compared to the average cell protein after a pulse of <~H leueine revealed that these polypeptides are derived from separate pools of precursor polypeptides. The pool sizes range from 2 p. cent to 25 p. cent of the amount of the corresponding polypeptide present in the cytochromes. The 32 000 molecular weight polypeptide of cytochrome band at least the 18 000 molecular weight polypeptide of cytochrome aa\(_3\) are mitochondrial translation products as well in the fungus Neurospora crassa as in the insect Locusta migratoria. So, despite the fact that the size of mitochondrial DNA and mitochondrial ribosomes is reduced in insects, the products have maintained their characteristics.},
  subject      = {Biochemie},
  language  = {en}
}
@article{JacklSebald1974,
  author    = {Jackl, G. and Sebald, Walter},
  title     = {Identification of two products of mitochondrial protein synthesis associated with oligomycin-sensitive ATPase from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82093},
  year      = {1974},
  abstract  = {no abstract available},
  subject      = {Physiologische Chemie},
  language  = {en}
}
@article{JacklSebald1975,
  author    = {Jackl, G. and Sebald, Walter},
  title     = {Identification of two products of mitochondrial protein synthesis associated with mitochondrial adenosine triphosphatase from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62812},
  year      = {1975},
  abstract  = {Soluble mitochondrial ATPase (F1) isolated from Neurospora crassa is resolved by dodecylsulfate- gel electrophoresis into five polypeptide bands with apparent molecular weights of 59000, 55000, 36000, 15000 and 12000. At least nine further polypeptides remain associated with ATPase after disintegration of mitochondria with Triton X-100 as shown by the analysis of an immunoprecipitate obtained with antiserum to F 1 A TPase. Two of the associated polypeptides with apparent molecular weights of 19000 and 11000 are translated on mitochondrial ribosomes, as demonstrated by incorporation in vivo of radioactive leueine in the presence of specific inhibitors of mitochondrial (chloramphenicol) and extramitochondrial ( cycloheximide) protein synthesis. The appearance of mitochondrial translation products in the immunoprecipitated A TPase complex is inhibited by' cycloheximide. The same applies for some of the extramitochondrial translation products in the presence of chloramphenicol. This suggests that both types of polypeptides are necessary for the assembly of the A TPase complex.},
  subject      = {Biochemie},
  language  = {en}
}
@article{Sebald1977,
  author    = {Sebald, Walter},
  title     = {Biogenesis of mitochondrial ATPase},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47362},
  year      = {1977},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{GrafSebald1978,
  author    = {Graf, T. and Sebald, Walter},
  title     = {The dicyclohexylcarbodiimide-binding protein of the mitochondrial ATPase complex from beef heart. Isolation and amino acid composition},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62806},
  year      = {1978},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{WeissSebald1978,
  author    = {Weiss, H. and Sebald, Walter},
  title     = {Purification of cytochrome oxidase from Neurospora crassa and other sources},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82082},
  year      = {1978},
  abstract  = {A chromatographic procedure 1 is described by means of which cytochrome oxidase has been purified from a variety of organisms including the fungus N eurospora crassa,2,3 the unicellular alga Po/ytoma mirum, 4 the insect Locusta migratoria ,5 the frog Xenopus muel/eri,4 and the mammal Rattus norwegicus. 4 This procedure can be used to equal effect for large-scale preparations, starting from grams of mitochondrial protein, or for small-scale preparations starting from milligrams. The cytochrome oxidase preparations from the different organisms are enzymically active. They show similar subunit compositions.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldWachterTzagoloff1979,
  author    = {Sebald, Walter and Wachter, E. and Tzagoloff, A.},
  title     = {Identification of amino acid substitutions in the dicyclohexylcarbodiimide-binding subunit of the mitochondrial ATPase complex from oligomycin-resistant mutants of Saccharomyces cerevisiae},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62770},
  year      = {1979},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{MichelWachterSebald1979,
  author    = {Michel, R. and Wachter, E. and Sebald, Walter},
  title     = {Synthesis of a larger precursor for the proteolipid subunit of the mitochondrial ATPase complex of Neurospora crassa in a cell-free wheat germ system},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62789},
  year      = {1979},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldGrafLukins1979,
  author    = {Sebald, Walter and Graf, T. and Lukins, H. B.},
  title     = {The dicyclohexylcarbodiimide-binding protein of the mitochondrial ATPase complex from Neurospora crassa and Saccharomyces cerevisiae. Identification and isolation},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62792},
  year      = {1979},
  abstract  = {Incubation of mitochondria from Neuraspara crassa and Saccharomyces cerevisiae with the radioactive ATPase inhibitor [14C]dicyclohexylcarbodiimide results in the irreversible and rather specific labelling of a low-molecular-weight polypeptide. This dicyclohexylcarbodiimide-binding protein is identical with the smallest subunit (Mr 8000) of the mitochondrial ATPase complex, and it occurs as oligomer, probably as hexamer, in the enzyme protein. The dicyclohexylcarbodiimide-binding protein is extracted from whole mitochondria with neutral chloroformjmethanol both in the free and in the inhibitor-modified form. In Neuraspara and yeast, this extraction is highly selective and the protein is obtained in homogeneaus form when the mitochondria have been prewashed with certain organic solvents. The bound dicyclohexylcarbodiimide Iabel is enriched in the purified protein up to 50-fold compared to whole mitochondria. Based on the amino acid analysis, the dicyclohexylcarbodiimide-binding protein from Neurospora and yeast consists of at least 81 and 76 residues, respectively. The content of hydrophobic residues is extremely high. Histidine and tryptophan are absent. The N-terminal ~mino acid is tyrosine in Neuraspara and formylmethionine in yeast.},
  subject      = {Biochemie},
  language  = {en}
}
@article{TzagoloffMacinoSebald1979,
  author    = {Tzagoloff, A. and Macino, G. and Sebald, Walter},
  title     = {Mitochondrial genes and translation products},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47408},
  year      = {1979},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldWernerWeiss1979,
  author    = {Sebald, Walter and Werner, S and Weiss, H},
  title     = {Biogenesis of mitochondrial membrane proteins in Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82055},
  year      = {1979},
  abstract  = {no abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldWild1979,
  author    = {Sebald, Walter and Wild, G.},
  title     = {Mitochondrial ATPase complex from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82065},
  year      = {1979},
  abstract  = {The A TPase eomplex has been isolated from mitoehondria of N eurospora crassa by immunologieal teehniques. The protein ean be obtained rapidly and qua ntitatively in high purity by miero- or large-seale immunopreeipitation. Immunopreeipitation has been applied to labeled and doubly labeled mitoehondrial proteins in order to investigate the number and moleeular weights of subunit polypeptides , the site of synthesis of subunit polypeptides, and the dieycIohexyIcarbodiimide-binding protein . The A TPase complex obtained by large-seale immunopreeipitation has been used as starting ma terial for the isolation of hydrophobie polypeptides.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldNeupertWeiss1979,
  author    = {Sebald, Walter and Neupert, W. and Weiss, H.},
  title     = {Preparation of Neurospora crassa mitochondria},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82070},
  year      = {1979},
  abstract  = {The fungus Neurospora crassa represents a eukaryotic cell with high biosynthetic activities. Cell mass doubles in 2-4 hr during expone ntial growth , even in simple salt media with sucrose as the sole carbon source. The microorgani sm forms a mycelium of long hyphae durlng vegetative growth . The mitochondria can be isolated under relatively gentle condi tions since a few breaks in the threadlike hyphae are sufficient to cause the outflow of the organelles. This article describes two methods for the physical disruption of the hyphae : (I) The cell s are opened in a grind mill between two rotating corundum di sks. This is a continuous and fast procedure and allows large- and small-scale preparations of mitochondria. (2) Hyphae are ground with sand in a mortar and pestle. This procedure can be applied to microscale preparations of mitochondria starting with minute amounts of cells. Other procedures for the isolation of Neurospora mitochondria after the physical di sruption or the enzymatic degradation of the cell wall have been described elsewhere},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeSebald1980,
  author    = {Hoppe, J. and Sebald, Walter},
  title     = {Amino acid sequence of the proteolipid subunit of the proton-translocating ATPase complex from the thermophilic bacterium PS-3},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62754},
  year      = {1980},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeSchairerSebald1980,
  author    = {Hoppe, J. and Schairer, H. U. and Sebald, Walter},
  title     = {The proteolipid of a mutant ATPase from Escherichia coli defective in H\(^+\)-conduction contains a glycine instead of the carbodiimide-reactive aspartyl residue},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62769},
  year      = {1980},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeSchairerSebald1980,
  author    = {Hoppe, J. and Schairer, HU and Sebald, Walter},
  title     = {Identification of amino-acid substitutions in the proteolipid subunit of the ATP synthase from dicyclohexylcarbodiimide-resistant mutants of Escherichia coli},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47374},
  year      = {1980},
  abstract  = {The amino acid sequence of the proteolipid subunit of the A TP synthase was analyzed in six mutant strains from Escherichia coli K 12, selected for their increased resistance towards the inhibitor N,N'-dicyclohexylcarbodiimide. All six inhibitor-resistant mutants were found to be altered at the same position of the proteolipid, namely at the isoleucine at residue 28. Two substitutions could be identified. In type I this residue was substituted by a valine resulting in a moderate decrease in sensitivity to dicyclohexylcarbodiimide. Type II contained a threonine residue at this position. Here a strong resistance was observed. These two amino acid substitutions did not influence functional properties of the ATPase complex. ATPase as well as A TP-dependent proton-translocating activities of mutant membranes were indistinguishable from the wild type. At elevated concentrations, dicyclohexylcarbodiimide still bound specifically to the aspartic acid at residue 61 of the mutant proteolipid as in the wild type, and thereby inhibited the activity of the ATPase complex. It is suggested that the residue 28 substituted in the resistant mutants interacts with dicyclohexylcarbodiimide during the reactions leading to the covalent attachment of the inhibitor to the aspartic acid at residue 61. This could indicate that these two residues are in close vicinity and would thus provide a first hint on the functional conformation of the proteolipid. Its polypeptide chain would have to fold back to bring together these two residues separated by a segment of 32 residues.},
  subject      = {Biochemie},
  language  = {en}
}
@article{vonJagowSebald1980,
  author    = {von Jagow, Gerhard and Sebald, Walter},
  title     = {b-Type cytochromes},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47383},
  year      = {1980},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldMachleidtWachter1980,
  author    = {Sebald, Walter and Machleidt, Werner and Wachter, Elmar},
  title     = {N,N'-dicyclohexylcarbodiimide binds specifically to a single glutamyl residue of the proteolipid subunit of the mitochondrial adenosinetriphosphatases from Neurospora crassa and Saccharomyces cerevisiae},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47394},
  year      = {1980},
  abstract  = {T~e N,N'-dicrclohexylcarbodiimide-binding proteolipid subumt of the mitochondrial adenosinetriphosphatases (ATP phosphohydrolase, EC 3.6.1.3) of Neurosporacrassa and Saccharomyces cerevisiae were purified from mitochondria incubated with the radioactively labeled inhibitor. The specifically labeled subunit was cleaved with cyanogen bromide and N-bromosuccinimide, and the resultant fragments were separated by gel chromatography in the presence of 80\% (vol/vol) formic acid. The N,N'-dicyclohexylcarbodiimide label was recovered in each organism exclusively in a 17-residue fragment. Further analysis by automated solid-phase Edman degrada.ti.on revealed tha~ the bound label was present at only one positIOn, correspondmg to a glutamyl residue. The NN'~ icyc~ohexyl~a~bodiiJ?1~de-'!l0dified glutamyl residue is the ~nly Id~ntIcal aCidic posItIon m both proteins and occurs in the middle of a hydrophobic sequence of about 25 residues.},
  subject      = {Dicyclohexylcarbodiimid},
  language  = {en}
}
@article{SchairerHoppeSebaldetal.1982,
  author    = {Schairer, H. U. and Hoppe, J. and Sebald, Walter and Friedl, P.},
  title     = {Topological and functional aspects of the proton conductor, F\(_0\), of the Escherichia coli ATP-synthase},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62721},
  year      = {1982},
  abstract  = {The isolated H\(^+\) conductor, F\(_0\) , of the Escherichia co1i ATP-synthase consists of three subunits, a, b, and c. H\(^+\) -permeable liposomes can be reconstit~ted with F\(_0\) and lipids; addition of F\(_1\)-ATPase reconstitutes a functional ATP-synthase. Mutants with altered or misslng F\(_0\) subunits are defective in H\(^+\) conduction. Thus, all three subunits are necessary for the expression of H\(^+\) conduction. The subunits a and b contain binding sites for F\(_1\)• Computer calculations, cross-links, membrane-permeating photo-reactive labels, and proteases were used to develop tentative structural models for the individual F\(_0\) subunits.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SebaldFriedlSchaireretal.1982,
  author    = {Sebald, Walter and Friedl, P. and Schairer, H. U. and Hoppe, J.},
  title     = {Structure and genetics of the H\(^+\)-conducting F\(_0\) portion of the ATP synthase},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62733},
  year      = {1982},
  abstract  = {The ATP synthase occurs in remarkably conserved form in procaryotic and eucaryotic cells. Thus, our present knowledge of ATP synthase is derived from sturlies of the enzyme from different organisms, each affering specific experimental possibilities. In recent tim es, research on the H\(^+\) -conducting F0 part of the ATP synthase has been greatly stimulated by two developments in the Escherichio coli system. Firstly, the purification and reconstitution of the whole ATP synthase as weil as the proton conductor Fa from E. coli have been achieved. These functionally active preparations are well defined in terms of subunit composition, similar to the thermophilic enzyme from PS-3 studied by Kagawa's group.u Secondly, the genetics and the molecular cloning of the genes of all the F\(_0\) subunits from E. coli yielded information on the function of subunit polypeptides and essential amino acid residues. Furthermore, the amino acid sequence of hydrophobic F\(_0\) subunits, which are difficult to analyze by protein-chemical techniques, could be derived from the nucleotide sequence of the genes. These achievements, which shall be briefly summarized in the next part of this communication, provide the framework to study specific aspects of the structure and function of the F\(_0\) subunits.},
  subject      = {Biochemie},
  language  = {en}
}
@article{ViebrockPerzSebald1982,
  author    = {Viebrock, A. and Perz, A. and Sebald, Walter},
  title     = {The imported preprotein of the proteolipid subunit of the mitochondrial ATP synthase from Neurospora crassa. Molecular cloning and sequencing of the mRNA},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62742},
  year      = {1982},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeFriedlSchaireretal.1983,
  author    = {Hoppe, J. and Friedl, P. and Schairer, H. U. and Sebald, Walter and Meyenburg, K. von and Jorgensen, B. B.},
  title     = {The topology of the proton translocating F\(_0\) component of the ATP synthase from E. coli K12: studies with proteases},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62718},
  year      = {1983},
  abstract  = {The accessibility of the three F\(_0\) subunits a, b and c from the Escherichia coli Kll A TP synthase to various proteases was studied in F\(_1\)-depleted inverted membrane vesicles. Subunit b was very sensitive to all applied proteases. Chymotrypsin produced a defined fragment of mol. wt. 1S 000 which remained tightly bound to the membrane. The cleavage site was located at the C-terminal region of subunit b. Larger amounts of proteases were necessary to attack subunit a (mol. wt. 30 000). There was no detectable deavage of subunit c. It is suggested that the major hydrophilic part of subunit b extends from the membrane into the cytoplasm and is in contact with the F\(_1\) sector. The F\(_1\) sector was found to afford some protection against proteolysis oftheb subunit in vitro andin vivo. Protease digestion bad no influence on the electro-impelled H\(^+\) conduction via F\(_0\) bot ATP-dependent H\(^+\) translocation could not be reconstituted upon binding of F\(_1\)• A possible role for subunit b as a linker between catalytic events on the F\(_1\) component and the proton pathway across the membrane is discussed.},
  subject      = {Biochemie},
  language  = {en}
}
@article{ViebrockPerzSebald1983,
  author    = {Viebrock, A and Perz, A and Sebald, Walter},
  title     = {Molecular cloning of middle-abundant mRNAs from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82033},
  year      = {1983},
  abstract  = {no abstract available},
  subject      = {Neurospora crassa},
  language  = {en}
}
@article{ArendsSebald1984,
  author    = {Arends, H. and Sebald, Walter},
  title     = {Nucleotide sequence of the cloned mRNA and gene of the ADP/ATP carrier from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62684},
  year      = {1984},
  abstract  = {A cDNA complementary to the mRNA of the ADPIATP carrier from Neurospora crassa was identified among ordered cDNA clones by hybridizing total polyadenylated RNA to pools of 96 cDNA recombinant plasmids and subsequent cellfree translation of hybridization-selected mRNA. Further carrier cDNAs were found by colony fdter hybridization at a frequency of 0.2-0.3\%. The gene of the carrier was cloned and isolated on a 4.6-kbp EcoRl fragment of total Neurospora DNA, and the start of the mRNA was determined by Sl nuclease mapping. From the nucleotide sequence of the cDNA and the genomic DNA, the primary structure of the gene, of the mRNA and of the ADP I ATP carrier protein could be deduced. The gene occurs in a single copy in the genome and related genes are absent. It contains two short introns, and a pyrimidine-rieb promoter region. The mRNA has a 46-bp 5 1 end and a 219-bp 3 1 end. There is an open reading frame coding for the 313 amino acid residues of the Neurospora carrier protein. The amino acid sequence is homologous in 148 positions with the established primary structure of the beef heart carrier.},
  subject      = {Biochemie},
  language  = {en}
}
@article{VeloursEsparzaHoppeetal.1984,
  author    = {Velours, J. and Esparza, M. and Hoppe, J. and Sebald, Walter and Guerin, B.},
  title     = {Amino acid sequence of a new mitochondrially synthesized proteolipid of the ATP synthase of Saccharomyces cerevisiae},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62695},
  year      = {1984},
  abstract  = {The purification and the amino acid sequence of a proteolipid translated on ribosomes in yeast mitochondria is reported. This protein, which is a subunit of the A TP synthase, was purified by extraction with chloroform/methanol (2/1) and subsequent chromatography on phosphocellulose and reverse phase h.p.l.c. A mol. wt. of 5500 was estimated by chromatography on Bio-Gel P-30 in 8011/o fonnie acid. The complete amino acid sequence of this protein was determined by automated solid phase Edman degradation of the whole protein and of fragments obtained after cleavage with cyanogen bromide. The sequence analysis indicates a length of 48 amino acid residues. The calculated mol. wt. of 5870 corresponds to the value found by gel chromatography. This polypeptide contains three basic residues and no negatively charged side chain. The three basic residues are clustered at the C terminus. The primary structure of this protein is in full agreement with the predicted amino acid sequence of the putative polypeptide encoded by the mitochondrial aap1 gene recently discovered in Saccharomyces cerevisiae. Moreover, this protein shows 5011/o homology with the amino acid sequence of a putative polypeptide encoded by an unidentified reading frame also discovered near the mitochondrial ATPase subunit 6 genein Aspergillus nidulans.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SchmidtWachterSebaldetal.1984,
  author    = {Schmidt, B. and Wachter, E. and Sebald, Walter and Neupert, W.},
  title     = {Processing peptidase of Neurospora mitochondria. Two-step cleavage of imported ATPase subunit 9},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62674},
  year      = {1984},
  abstract  = {Subunit 9 (dicyclohexylcarbod{\"u}mide binding protein, 'proteolipid') of the mitochondrial F 1F0-ATPase is a nuclearly coded protein in Neurospora crassa. lt is synthesized on free cytoplasmic ribosomes as a larger precursor with an NH2-terminal peptide extension. The peptide extension is cleaved ofT after transport of the protein into the mitochondria. A processing activity referred to as processing peptidase that cleaves the precursor to subunit 9 and other mitochondrial proteins is described and characterized using a cell-free system. Precursor synthesized in vitro was incubated with extracts of mitochondria. Processing peptidase required Mn2 + for its activity. Localization studies suggested that it is a soluble component of the mitochondrial matrix. The precursor was cleaved in two sequential steps via an intermediate-sized polypeptide. The intermediate form in the processing of subunit 9 was also seen in vivo and upon import of the precursor into isolated mitochondria in vitro. The two dcavage sites in the precursor molecule were determined. The data indicate that: {a) the correct NH2-terminus of the mature protein was generated, (b) the NH2-terminal amino acid of the intermediate-sized polypeptide is isoleueine in position -31. The cleavage sites show similarity ofprimary structure. It is concluded that processing peptidase removes the peptide extension from the precursor to subunit 9 (and probably other precursors) after translocation of these polypeptides (or the NHrterminal part of these polypeptides) into the matrix space of mitochondria.},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeSebald1984,
  author    = {Hoppe, J. and Sebald, Walter},
  title     = {The proton conducting F0-part of bacterial ATP synthases},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-82019},
  year      = {1984},
  abstract  = {No abstract available},
  subject      = {Biologie},
  language  = {en}
}
@article{SebaldArendsMcCarthy1984,
  author    = {Sebald, Walter and Arends, Hermann and McCarthy, John E. G.},
  title     = {Isolation and manipulation of genes coding for energy-transducing enzymes from Neurospora crassa and Escherichia coli},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-86768},
  year      = {1984},
  abstract  = {No abstract available.},
  subject      = {Escherichia coli},
  language  = {en}
}
@article{HarnischWeissSebald1985,
  author    = {Harnisch, U. and Weiss, H. and Sebald, Walter},
  title     = {The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora, determined by cDNA and gene sequencing},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62631},
  year      = {1985},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{GabelliniHarnischMcCarthyetal.1985,
  author    = {Gabellini, N. and Harnisch, U. and McCarthy, J. E. and Hauska, G. and Sebald, Walter},
  title     = {Cloning and expression of the fbc operon encoding the FeS protein, cytochrome b and cytochrome c\(_1\) from the Rhodopseudomonas sphaeroides b/c\(_1\) complex},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62642},
  year      = {1985},
  abstract  = {The gene for the FeS protein of the Rhodopseudomonas sphaeroides b/c1 complex was identified by means of crosshybridization with a segment of the gene encoding the corresponding FeS protein of Neurospora crassa. Plasmids (pRSF1-14) containing the cross-hybridizing region, covering in total 13.5 kb of chromosomal DNA, were expressed in vitro in a homologous system. One RSF plasmid directed the synthesis of all three main polypeptides of the R. sphaeroides blc1 complex: the FeS protein, cytochrome b and cytochrome c1• The FeS protein and cytochrome c1 were apparently synthesized as precursor fonns. None of the pRSF plasmids directed the synthesis of the 10-kd polypeptide found in b/c1 complex preparations. Partial sequencing of the cloned region was performed. Several sites of strong homology between R. sphaeroides and eukaryotic polypeptides of the b/c1 complex were identified. The genes encode the three b/c1 polypeptides in the order: (5') FeS protein, cytochrome b, cytochrome c1• The three genes are transcribed to give a polycistronic mRNA of 2.9 kb. This transcriptional unit has been designated the jbc operon; its coding capacity corresponds to the size of the polycistronic mRNA assuming that only the genes for the FeS protein (jbcF), cytochrome b (jbcß) and cytochrome c1 (jbcC) are present. This could indicate that these three subunits constitute the minimal catalytic unit of the b/c1 complex from photosynthetic membranes.},
  subject      = {Biochemie},
  language  = {en}
}
@article{McCarthySchairerSebald1985,
  author    = {McCarthy, J. E. and Schairer, H. U. and Sebald, Walter},
  title     = {Translational initiation frequency of atp genes from Escherichia coli: identification of an intercistronic sequence that enhances translation},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62657},
  year      = {1985},
  abstract  = {The c, b and {\"o} subunit genes of the Escherichia coli atp operon were cloned individually in an expression vector between the tac fusion promoter and the galK gene. The relative rates of subunit synthesis directed by the cloned genes were similar in vitro andin vivo and compared favourably with the subunit stoichiometry of the assembled proton-translocating A TP synthase of E. coli in vivo. The rate of synthesis of subunit c was at least six times that of subunit b and 18 times that of subunit {\"o}. Progressive shortening of the long intercistronic sequence lying upstream of the subunit c gene showed that maximal expression of this gene is dependent upon the presence of a sequence stretching > 20 bp upstream of the Shine-Dalgarno site. This sequence thus acts to enhance the rate of translational initiation. The possibility that similar sequences might perform the same function in other operons of E. coli and bacteriophage A is also discussed. Translation of the subunit b cistron is partially coupled to translation of the preceding subunit c cistron. In conclusion, the expression of all the atp operon genes could be adjusted to accommodate the subunit requirements of A TP synthase assembly primarily by means of mechanisms which control the efficiency of translational initiation and re-initiation at the respective cistron start codons.},
  subject      = {Biochemie},
  language  = {en}
}
@article{LindenmaierDittmarHauseretal.1985,
  author    = {Lindenmaier, W. and Dittmar, K. E. and Hauser, H. and Necker, A. and Sebald, Walter},
  title     = {Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62662},
  year      = {1985},
  abstract  = {A method has been developed that allows the isolation of genomic clones from a cosmid library by homologaus recombination in vivo. This method was used to isolate a human genomic interleukin 2 (IL2) gene. The genomic cosmid library was packaged in vivo into A. phage particles. A recombination-proficient host strain carrying IL2 cDNA sequences in a non-homologaus plasmid vector was infected by the packaged cosmid library. After in vivo packaging and reinfection, recombinants carrying the antibiotic resistance genes of both vectors were selected. From a recombinant cosmid clone the chromosomal IL2 genewas restored. After DNA mediated gene transfer into mouse Ltk- cells human IL2 was expressed constitutively.},
  subject      = {Biochemie},
  language  = {en}
}
@article{WeichSebaldSchaireretal.1986,
  author    = {Weich, H. A. and Sebald, Walter and Schairer, H. U. and Hoppe, J.},
  title     = {The human osteosarcoma cell line U-2 OS expresses a 3.8 kilobase mRNA which codes for the sequence of the PDGF-B chain},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62588},
  year      = {1986},
  abstract  = {A cDNA clone of about 2500 basepairswas prepared from the human osteosarcoma cellline U-2 OS by hybridizing with a v-sis probe. Sequence analysis showed that this cDNA contains the coding region for the PDGF-B chain. Here we report that the mitogen secreted by these osteosarcoma cells contains the PDGF-B chain and is probably a homodimer of two B-chains.},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeGattiWeberetal.1986,
  author    = {Hoppe, J. and Gatti, D. and Weber, H. and Sebald, Walter},
  title     = {Labeling of individual amino acid residues in the membrane-embedded F\(_0\) part of the F\(_1\) F\(_0\) ATP synthase from Neurospora crassa. Influence of oligomycin and dicyclohexylcarbodiimide},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62598},
  year      = {1986},
  abstract  = {Three F0 subunits and the F\(_1\) subunit P of the ATP synthase from Neurospora crassa were labeled with the lipophilic photoactivatable reagent 3-(trifluoromethyl)-3-(m-[\(^{125}\)I]iodophenyl)diazirine ([\(^{125}\)I]TID). In the proteolipid subunit which was the most heavily labeled polypeptide labeling was confmed to five residues at the NH2-terminus and five residues at the C-terminus ofthe protein. Labeling occurred at similar positions compared with the homologaus protein (subunit c) in the ATP synthase from Escherichia coli, indicating a similar structure of the proteolipid subunits in their respective organisms. The inhibitors oligomycin and dicyclohexylcarbodiimide did not change the pattern of accessible surface residues in the proteolipid, suggesting that neither inhibitor induces gross conformational changes. However, in the presence of oligomycin, the extent oflabeling in some residues was reduced. Apparently, these residues provide part of the binding site for the inhibitor. After reaction with dicyclohexylcarbodiimide an additional labeled amino acid was found at position 65 corresponding to the invariant carbod{\"u}mide-binding glutamic acid. These results and previous observations indicate that the carboxyl side chain of Glu-65 is located at the protein-lipid interphase. The idea is discussed that proton translocation occurs at the interphase between different types if F\(_0\) subunits. Dicyclohexylcarbodiimide or oligomycin might disturb this essential interaction between the F\(_0\) subunits.},
  subject      = {Biochemie},
  language  = {en}
}
@article{HoppeSebald1986,
  author    = {Hoppe, J. and Sebald, Walter},
  title     = {Topological studies suggest that the pathway of the protons through F\(_0\) is provided by amino acid residues accessible from the lipid phase},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62602},
  year      = {1986},
  abstract  = {The structure of the F0 part of ATP synthases from E. coli and Neurospora crassa was analyzed by hydrophobic surface labeling with [125I]TID. In the E. co/i F0 all three subunits were freely accessible to the reagent, suggesting that these subunits are independently integrated in the membrane. Labeted amino acid residues were identified by Edman degradation of the dicyclohexylcarbodiimide binding (DCCD) proteins from E. coli and Neurospora crassa. The very similar patterns obtained with the two homologaus proteins suggested the existence of tightly packed cx-helices. The oligomeric structure of the DCCD binding protein appeared to be very rigid since little, if any, change in the labeling patternwas observed upon addition of oligomycin or DCCD to membranes from Neurospora crassa. When membrancs were pretrcated with DCCD prior to the reaction with [125I]TID an additionally labeled amino acid appeared at the position of Glu·65 which binds DCCD covalently, indicating the Jocation of this inhibitor on the outside of the oligomer. It is suggested that proton conduction occurs at the surface of the oligomer of the DCCD binding protein. Possibly this oligomer rotates against the subunit a or b and thus enables proton translocation. Conserved residues in subunit a, probably located in the Iipid bilayer, might participate in the pro· ton translocation mechanism.},
  subject      = {Biochemie},
  language  = {en}
}
@article{GabelliniSebald1986,
  author    = {Gabellini, N. and Sebald, Walter},
  title     = {Nucleotide sequence and transcription of the fbc operon from Rhodopseudomonas sphaeroides. Evaluation of the deduced amino acid sequences of the FeS protein, cytochrome b and cytochrome c\(_1\)},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62615},
  year      = {1986},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{McCarthySebaldGrossetal.1986,
  author    = {McCarthy, J. E. and Sebald, Walter and Gross, G. and Lammers, R.},
  title     = {Enhancement of translational efficiency by the Escherichia coli atpE translational initiation region: its fusion with two human genes},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62626},
  year      = {1986},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{KleenePfannerPfalleretal.1987,
  author    = {Kleene, R. and Pfanner, N. and Pfaller, R. and Link, T. A. and Sebald, Walter and Neupert, W. and Tropschug, M.},
  title     = {Mitochondrial porin of Neurospora crassa: cDNA cloning, in vitro expression and import into mitochondria},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62566},
  year      = {1987},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{RoemischTropschugSebaldetal.1987,
  author    = {R{\"o}misch, J. and Tropschug, M. and Sebald, Walter and Weiss, H.},
  title     = {The primary structure of cytochrome c\(_1\) from Neurospora crassa},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62578},
  year      = {1987},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{WeigelMeyerSebald1989,
  author    = {Weigel, U. and Meyer, M. and Sebald, Walter},
  title     = {Mutant proteins of human interleukin 2. Renaturation yield, proliferative activity and receptor binding},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62543},
  year      = {1989},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{FlueggeFischerGrossetal.1989,
  author    = {Fl{\"u}gge, U. I. and Fischer, K. and Gross, A. and Sebald, Walter and Lottspeich, F. and Eckerskorn, C.},
  title     = {The triose phosphate-3-phosphoglycerate-phosphate translocator from spinach chloroplasts: nucleotide sequence of a full-length cDNA clone and import of the in vitro synthesized precursor protein into chloroplasts},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62559},
  year      = {1989},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{MerzFliednerLehrnbecheretal.1990,
  author    = {Merz, H. and Fliedner, A. and Lehrnbecher, T. and Sebald, Walter and M{\"u}ller-Hermelink, H. K. and Feller, A. C.},
  title     = {Cytokine expression in B-cell non-Hodgkin lymphomas},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62539},
  year      = {1990},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{LehrnbecherMerzSebaldetal.1991,
  author    = {Lehrnbecher, T. and Merz, H. and Sebald, Walter and Poot, M.},
  title     = {Interleukin 4 drives phytohemagglutinin-activated T cells through several cell cycles: no synergism between interleukin 2 and interleukin 4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62491},
  year      = {1991},
  abstract  = {Cell kinetic studies of T cells stimulated with the interleukin 2 (11-2), D-4, or both lymphokines were performed with conventional [3H] thymidine incorporation and with the bivariate BrdU/Hoechst technique. 11-2 and 11-4 are able to drive phytohemagglutininactivated T cells through more than one cell cycle. Neither synergistic nor inhibitory efl'ect on T -cell proliferationwas seen for the stimulation with both 11-2 and 11-4 as compared with the effect ofll-2 alone. The quantitative data ofthe cell cycle distribution ofphytohemagglutininactivated T cells suggestthat the population ofll-4-responsive cells is at least an overlapping population, if not a real subset of the ·population of the 11-2-responsive cells.},
  subject      = {Biochemie},
  language  = {en}
}
@article{MerzFliednerOrschescheketal.1991,
  author    = {Merz, H. and Fliedner, A. and Orscheschek, K. and Binder, T. and Sebald, Walter and M{\"u}ller-Hermelink, H. K. and Feller, A. C.},
  title     = {Cytokine expression in T-cell lymphomas and Hodgkin's disease. Its possible implication in autocrine or paracrine production as a potential basis for neoplastic growth},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62483},
  year      = {1991},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{KruseLehrnbecherSebald1991,
  author    = {Kruse, N. and Lehrnbecher, T. and Sebald, Walter},
  title     = {Site-directed mutagenesis reveals the importance of disulfide bridges and aromatic residues for structure and proliferative activity of human interleukin-4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62505},
  year      = {1991},
  abstract  = {Mutant proteins (muteins) of human lnterleukin-4 (llA) were constructed by means of in vitro mutagenesis. The muteins were expressed in E. co/1, submitted to a renaturation and purification protocol and analysed for biological activity. Exchange of the cysteines at either position 46 or 99 which form one of the three disulfide bridges resulted. in a nearly co•mplete loss · of biological actiyity and an unstable protein. The exchange of tyrosine 124 also inactivated the protein, while a mutation of tyrosine 56 left some residual activity. Exchange of the other four cysteines or of · the single tryptophane had smaller etTects.},
  subject      = {Biochemie},
  language  = {en}
}
@article{KruseTonySebald1992,
  author    = {Kruse, N. and Tony, H. P. and Sebald, Walter},
  title     = {Conversion of human interleukin-4 into a high affinity antagonist by a single amino acid replacement},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62469},
  year      = {1992},
  abstract  = {lnterleukin-4 (IL-4) represents a prototypic lymphokine (for a recent review see Paul, 1991). It promotes differentiation of B-cells and the proliferation of T- and B-cell, and other cell types of the lymphoid system. An antagonist of human IL-4 was discovered during the studies presented here after Tyr124 of the recombinant proteinbad been substituted by an aspartic acid residue. This IL-4 variant, Y124D, bound with high affinity to the IL-4 receptor (K\(_D\) = 310 pM), but retained no detectable proliferative activity for T -<:ells and inhibited IL-4-dependent T -cell proliferation competitively (K\(_i\) = 620 pM). The loss of efficacy in variant Y124D was estimated to be > 100-fold on the basis of a weak partial agonist activity for the very sensitive induction of CD23 positive B-cells. The subsitution of Tyr124 by either phenylalanine, histidine, asparagine, Iysine or glycine resulted in partial agonist variants with unaltered receptor binding atTmity and relatively small deficiencies in efficacy. These results demoostrate that high affinity binding and signal generation can be uncoupled efticiently in a Iigand of a receptor betonging to the recently identified hematopoietin receptor family. In addition we show for the first time, that a powerful antagonist acting on the IL-4 receptor system can be derived from the IL-4 protein.},
  subject      = {Biochemie},
  language  = {en}
}
@article{DummerPosseckertNestleetal.1992,
  author    = {Dummer, R. and Posseckert, G. and Nestle, F. and Witzgall, R. and Burger, M. and Becker, J. C. and Sch{\"a}fer, E. and Wiede, J. and Sebald, Walter and Burg, G.},
  title     = {Soluble interleukin-2 receptors inhibit interleukin 2-dependent proliferation and cytotoxicity: explanation for diminished natural killer cell activity in cutaneous T-cell lymphomas in vivo?},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62473},
  year      = {1992},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{DuschlJahnBertlingetal.1992,
  author    = {Duschl, Albert and Jahn, Ute and Bertling, Claudia and Sebald, Walter},
  title     = {A comparison of assays for the response of primary human T-cells upon stimulation with interleukin-2, interleukin-4 and interleukin-7},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-86750},
  year      = {1992},
  abstract  = {The most commonly used assay to quantitate the response of peripheral T~cells upon stimulation with growth factors is determination of incorporated (JH]TdR. We compared thls test to three other methods: 1. direct countlog of cells with a Coulter type counter as reference assay, 2. a colorimetric assay using the tetrazolium dye 3-[ 4,S-dimethylthiazol-l-yl]-2,5diphenyl tetrazolium (MTT), which is a cheap and increasingly popular non-radioactive method and 3. incorporation of the thymidine analog 5-bromo-2'-deoxyuridine detection with a monoclonal antibody on cytospins. Primary human PHA-blasts from >30 healthy individuals were stimulated with IL-2, IL-4 aod IL-7 and assayed with up to four different methods. We discuss the advantages and disadvantages of the assays used and tbe effects of differences between cell preparations. We observed no significant variations between individuals for the dose dependence, but the relative emctency of IL4 compared to IL-2 and IL-7 was variable. This was probably due to the slower response observed upon stimulation with this factor.},
  subject      = {T-Lymphozyt},
  language  = {en}
}
@article{KruseShenArnoldetal.1993,
  author    = {Kruse, N. and Shen, B. J. and Arnold, S. and Tony, H. P. and M{\"u}ller, T. and Sebald, Walter},
  title     = {Two distinct functional sites of human interleukin 4 are identified by variants impaired in either receptor binding or receptor activation},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62451},
  year      = {1993},
  abstract  = {Interleukin 4 (IL-4) exerts a decisive role in the coord.ination of proteelive immune responses against parasites, particularly helminths. A disregulation of ll.r4 function is possibly involved in the genesis of allergic disease states. The search for important amino acid residues in human ll.r4 by mutational analysis of charged invariant amino acid positions identified two distinct functional sites in the 4-helix-bundle protein. Site 1 was marked by amino acid substitutions of the glutamic acid at position 9 in helix A and arginine at position 88 in helix C. Exchanges at both positions led to IL-4 variants deficient in binding to the extracellular domain of the ll.r4 receptor (IL-4ReJ. In parallel, up to 1000-fold increased concentrations of this type of variant were required to induce T -cell proliferation and B-eeil CD23 expression. Site 2 was marked by amino acid exchanges in helix D at positions 121, 124 and 125 (arginine, tyrosine and serine respectively in the wild-type).ß.A variants affected at site 2 exhibited partial agonist activity during T -cell proliferation; however, they still bound with high affinity to IL-4Rex. [The generation of an IL-4 antagonist by replacing tyrosine 124 with aspartic acid has been described before by Kruse et al. (1992) (EMBO }., 11, 3237-3244)]. These findings indicate that IL-4 functions by bind.ing IL-4Rex via site 1 which is constituted by residues on helices A and C. They further suggest that the association of a second, still undetined receptor protein with site 2 in helix D activates the receptor system and generates a transmembrane signal.},
  subject      = {Biochemie},
  language  = {en}
}
@article{KueblerReutherKirchneretal.1993,
  author    = {K{\"u}bler, Norbert and Reuther, J{\"u}rgen and Kirchner, Thomas and Priessnitz, Bernd and Sebald, Walter},
  title     = {Osteoinductive, morphologic, and biomechanical properties of autolyzed, antigen-extracted, allogeneic human bone},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-86715},
  year      = {1993},
  abstract  = {Autolyzed, antigen-extracted, allogeneic (AAA) bone was prepared from human cortical bone and its morphologic, biomechanical, and osteoinductive properlies were compared with untreated (frozen) as well as lyophilized human bone. Scanning electron microscopy revealed removal of inprganic calcium phosphates and persistence of shrunken collagen fibrils on the surface of AAA bone matrix. Biomechanical testing of differently prepared bone samples showed that lyophilization increased both the modulus of elasticity (P < .00001) and the compressive strength (P < .00001 ). Depending on the depth of decalcification in the preparation of AAA bone, both measured values decreased in rehydrated AAA bone compared with untreated bone {P < .00001 ). Completely demineralized and rehydrated AAA bone was soft, flexible, and showed very little compressive strength. Differences in biomechanical behavior between samples drilled longitudinally or perpendicularly to the diaphyseal bone axis were observed. Xenogeneic human bone samples were implanted in muscle pouches of Sprague-Dawley rats for 6 weeks. AAA bone implants showed chondrogenesis and osteogenesis in 50\% of the cases, while untreated or lyophilized bone implants induced no new cartilage or bone formation. As decalcification exposed xenogeneic organic matrix components, AAA bone implants provoked the highest inflammatory reaction. When AAA bone samples were implanted in immunosuppressed rats, the inflammatory reaction was suppressed and 94o/o of the implants showed endochondral bone formation. The chondroinductivity of the bone samples also was tested in vitro using neonatal rat muscle tissue to avoid interference with inflammatory cells and secreted cytokines. In this assay, 68°/o of AAA bone samples induced chondroneogenesis, while untreated as weil as lyophilized bone samples failed to induce any cartilage formation. The results clearly dernonstrafe that AAA bone has osteoinductive properties. Biomechanical stability of AAA bone implants depends on the degree of demineralization. Thus, they can be prepared in an appropriate manner for different indications in oral and maxillofacial surgery.},
  subject      = {Mund-Kiefer-Gesichts-Chirurgie},
  language  = {en}
}
@article{MuellerDieckmannSebaldetal.1994,
  author    = {M{\"u}ller, T. and Dieckmann, T. and Sebald, Walter and Oschkinat, H.},
  title     = {Aspects of receptor binding and signalling of interleukin-4 investigated by site-directed mutagenesis and NMR spectroscopy},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62444},
  year      = {1994},
  abstract  = {Cytokines are hormones that carry information from ceJI to ceH. This information is read from their surface upon binding to transmembrane receptors and by the subsequent initiation of receptor oligomerization. An inftuence on this process through mutagenesis on the hormone surface is highly desirab)e for medical reasons. However, an understanding of hormone-receptor interactions requires insight into the structural changes introduced by the mutations. In this line structural studies on human TL-4 and the medically important IL-4 antagonists YI24D and Y124G are presented. The site a.round YI24 is an important epitope responsible for the a.bility of 11-4 t.o ca.use a signal in the target cells. It is shown that the local main-chain structure around residue 124 in the variants remains unchanged. A strategy is presented here which allows the study of these types of proteins and their variants by NMR which does not require carbon Iabeiied sa.mples.},
  subject      = {Biochemie},
  language  = {en}
}
@article{MuellerSebaldOschkinat1994,
  author    = {M{\"u}ller, T. and Sebald, Walter and Oschkinat, H.},
  title     = {Antagonist design through forced electrostatic mismatch},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62408},
  year      = {1994},
  abstract  = {No abstract available},
  subject      = {Biochemie},
  language  = {en}
}
@article{ReuschArnoldHeusseretal.1994,
  author    = {Reusch, P. and Arnold, S. and Heusser, C. and Wagner, K. and Weston, B. and Sebald, Walter},
  title     = {Neutralizing monoclonal antibodies define two different functional sites in human interleukin-4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62418},
  year      = {1994},
  abstract  = {Human interleukin-4 (IL-4) is a small four-helix-bundle protein which is essential for organizing defense reactions against macroparasites, in particular helminths. Human IL-4 also appears to exert a pathophysiological role during various IgE-mediated allergic diseases. Seven different monoclonal antibodies neutralizing the activity of human IL-4 were studied in order to identify functionally important epitopes. A collection of 41 purified IL-4 variants was used to analyse how defined amino acid replacements affect binding affinity for each individual mAb. Specific amino acid positions could be assigned to four different epitopes. mAbs recognizing epitopes on helix A and/or C interfered with IL-4 receptor binding and thus inhibited IL-4 function. However, other mAbs also inhibiting IL-4 function recognized an epitope on helix D of IL-4 and did not inhibit IL-4 binding to the receptor protein. One mAb, recognizing N-terminal and C-terminal residues, partially competed for binding to the receptor. The results of these mAb epitope analyses confirm and extend previous data on the functional consequences of the amino acid replacements which showed that amino acid residues in helices A and C of IL-4 provide a binding site for the cloned IL-4 receptor and that a signalling site in helix D interacts with a further receptor protein.},
  subject      = {Biochemie},
  language  = {en}
}
@article{DemchukMuellerOschkinatetal.1994,
  author    = {Demchuk, E. and Mueller, T. and Oschkinat, H. and Sebald, Walter and Wade, R. C.},
  title     = {Receptor binding properties of four-helix-bundle growth factors deduced from electrostatic analysis},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62424},
  year      = {1994},
  abstract  = {Hormones of the hematopoietin class mediate signal transduction by binding to specific transmembrane receptors. Structural data show that the human growth hormone (hGH) forms a complex with a homodimeric receptor and that hGH is a member of a class of hematopoietins possessing an antiparallel 4-a-helix bundle fold. Mutagenesis experiments suggest that electrostatic interactions may have an important influence on hormonereceptor recognition. In order to examine the specificity of hormone-receptor complexation, an analysis was made of the electrostatic potentials of hGH, interleukin-2 (IL-2), interleukin-4 (IL-4), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and the hGH and IL-4 receptors. The binding surfaces of hGH and its receptor, and of IL-4 and its receptor, show complementary electrostatic potentials. The potentials of the hGH and its receptor display approximately 2-fold rotational symmetry because the receptor subunits are identical. In contrast, the potentials of GM-CSF and IL-2 Iack such symmetry, consistent with their known high affinity for hetero-oligomeric receptors. Analysis of the electrostatic potentials supports a recently proposed hetero-oligomeric model for a high-affinity IL-4 receptor and suggests a possible new receptor binding mode for G-CSF; it also provides valuable information for guiding structural and mutagenesis studies of signal-transducing proteins and their receptors.},
  subject      = {Biochemie},
  language  = {en}
}
@article{LehrnbecherPootOrschescheketal.1994,
  author    = {Lehrnbecher, T. and Poot, M. and Orscheschek, K. and Sebald, Walter and Feller, A. C. and Merz, H.},
  title     = {Interleukin 7 as interleukin 9 drives phytohemagglutinin-activated T cells through several cell cycles; no synergism between interleukin 7, interleukin 9 and interleukin 4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62438},
  year      = {1994},
  abstract  = {The effects of the interlenkins IL-7 and IL-9 on cell cycle progression were investigated by conventional [3H]thymidine incorporation and by the bivariate BrdU/Hoechst technique. 8oth IL· 7 and IL-9 drive phytohemagglutinin-activated T cells through more than one cell cycle, but IL-7 wasmorepotent on cell cycle progression than IL-9. Neither synergistic nor inhibitory effects were seen between various combinations of the lymphokines IL-7, IL-9 and IL-4 compared to each lymphokine alone. When T cells are activated with phytohemagglutinin for 3 days, all or most IL-4 responsive cells respond to IL-7 as weil, whereas only a part of IL-7 responders are IL-4 responders. In contrast, when T cells are activated with phytohemagglutinin for 7 days, the quantitative data of the cell cycle distribution soggest that the population of IL-7 responders is at least an overlapping, if not a real subset of the population of the IL-4 responders.},
  subject      = {Biochemie},
  language  = {en}
}
@article{KueblerReutherKirchneretal.1994,
  author    = {K{\"u}bler, N. and Reuther, J. and Kirchner, T. and Pfaff, M. and M{\"u}ller-Hermelink, H. K. and Albert, R. and Sebald, Walter},
  title     = {IgG monoclonal antibodies that inhibit osteoinductivity of human bone matrix-derived proteins (hBMP/NCP)},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62388},
  year      = {1994},
  abstract  = {Monoclonal hBMP/NCP (human bone morphogenetic protein anrl associaterl noncollagenous proteins) antiborlies of the lgG class were prorlucerl. In vitro, 12 of 19 hBMP/NCP antiborlies showerl functional inhibition of hBMP/ NCP-induced chondroneogenesis in a neonatal muscle tissue assay. Inducing factors were characterized by their inhibiting antibodies with immunoblotting. Several peptide factors seem to be involved in the cascade of inducerl chondro- and osteogenesis.},
  subject      = {Biochemie},
  language  = {en}
}
@article{TonyShenReuschetal.1994,
  author    = {Tony, H. P. and Shen, B. J. and Reusch, P. and Sebald, Walter},
  title     = {Design of human interleukin-4 antagonists inhibiting interleukin-4-dependent and interleukin-13-dependent responses in T-cells and B-cells with high efficiency},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-62394},
  year      = {1994},
  abstract  = {Human interleukin-4 possesses two distinct sites for receptor activation. A signaHing site, comprising residues near the C-terminus on helix D, determines the efficacy of interleukin-4 signal transduction without affecting the binding to the interleukin-4 receptor a subunit. A complete antagonist and a series of low-efficacy agonist variants of human interleukin-4 could be generated by introducing combinations of two or three negatively charged aspartic acid residues in this site at positions 121, 124, and 125. One of the double variants, designated [R121D,Y124D]interleukin-4, with replacements of b{\"o}th Arg121 and Tyr124 by aspartic acid residues was completely inactive in all analysed cellular responses. The loss of efficacy in [R121D,Y124D]interleukin-4 is estimated to be larger than 2000-fold. Variant [R121D,Y124D]interleukin-4 was also a perfect antagonist for inhibition of interleukin-13-dependent responses in B-cells and the TF-1 cellline with a K\(_i\) value of approximately 100 pM. In addition, inhibition of both interleukin-4-induced and interleuk.in-13- induced responses could be obtained by monoclonal antibody X2/45 raised against interleukin-4Rm the extracellular domain of the interleuk.in-4 receptor a subunit. These results indicate that efficient interleukin-4 antagonists can be designed on the basis of a sequential two-step activation model. In addition, the experiments indicate the functional participation of the interleukin-4 receptor a subunit in the interleukin-13 receptor system.},
  subject      = {Biochemie},
  language  = {en}
}
@article{SeherNickelMuelleretal.2011,
  author    = {Seher, Axel and Nickel, Joachim and Mueller, Thomas D. and Kneitz, Susanne and Gebhardt, Susanne and Meyer ter Vehn, Tobias and Schlunck, Guenther and Sebald, Walter},
  title     = {Gene expression profiling of connective tissue growth factor (CTGF) stimulated primary human tenon fibroblasts reveals an inflammatory and wound healing response in vitro},
  series = {Molecular Vision},
  volume    = {17},
  journal   = {Molecular Vision},
  number    = {08. Okt},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-140189},
  pages     = {53-62},
  year      = {2011},
  abstract  = {Purpose: The biologic relevance of human connective tissue growth factor (hCTGF) for primary human tenon fibroblasts (HTFs) was investigated by RNA expression profiling using affymetrix (TM) oligonucleotide array technology to identify genes that are regulated by hCTGF. Methods: Recombinant hCTGF was expressed in HEK293T cells and purified by affinity and gel chromatography. Specificity and biologic activity of hCTGF was confirmed by biosensor interaction analysis and proliferation assays. For RNA expression profiling HTFs were stimulated with hCTGF for 48h and analyzed using affymetrix (TM) oligonucleotide array technology. Results were validated by real time RT-PCR. Results: hCTGF induces various groups of genes responsible for a wound healing and inflammatory response in HTFs. A new subset of CTGF inducible inflammatory genes was discovered (e.g., chemokine [C-X-C motif] ligand 1 [CXCL1], chemokine [C-X-C motif] ligand 6 [CXCL6], interleukin 6 [IL6], and interleukin 8 [IL8]). We also identified genes that can transmit the known biologic functions initiated by CTGF such as proliferation and extracellular matrix remodelling. Of special interest is a group of genes, e.g., osteoglycin (OGN) and osteomodulin (OMD), which are known to play a key role in osteoblast biology. Conclusions: This study specifies the important role of hCTGF for primary tenon fibroblast function. The RNA expression profile yields new insights into the relevance of hCTGF in influencing biologic processes like wound healing, inflammation, proliferation, and extracellular matrix remodelling in vitro via transcriptional regulation of specific genes. The results suggest that CTGF potentially acts as a modulating factor in inflammatory and wound healing response in fibroblasts of the human eye.},
  language  = {en}
}
@article{MuellerFiebigWeidaueretal.2013,
  author    = {Mueller, Thomas D. and Fiebig, Juliane E. and Weidauer, Stella E. and Qiu, Li-Yan and Bauer, Markus and Schmieder, Peter and Beerbaum, Monika and Zhang, Jin-Li and Oschkinat, Hartmut and Sebald, Walter},
  title     = {The Clip-Segment of the von Willebrand Domain 1 of the BMP Modulator Protein Crossveinless 2 Is Preformed},
  series = {Molecules},
  journal   = {Molecules},
  doi       = {10.3390/molecules181011658},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-97196},
  year      = {2013},
  abstract  = {Bone Morphogenetic Proteins (BMPs) are secreted protein hormones that act as morphogens and exert essential roles during embryonic development of tissues and organs. Signaling by BMPs occurs via hetero-oligomerization of two types of serine/threonine kinase transmembrane receptors. Due to the small number of available receptors for a large number of BMP ligands ligand-receptor promiscuity presents an evident problem requiring additional regulatory mechanisms for ligand-specific signaling. Such additional regulation is achieved through a plethora of extracellular antagonists, among them members of the Chordin superfamily, that modulate BMP signaling activity by binding. The key-element in Chordin-related antagonists for interacting with BMPs is the von Willebrand type C (VWC) module, which is a small domain of about 50 to 60 residues occurring in many different proteins. Although a structure of the VWC domain of the Chordin-member Crossveinless 2 (CV2) bound to BMP-2 has been determined by X-ray crystallography, the molecular mechanism by which the VWC domain binds BMPs has remained unclear. Here we present the NMR structure of the Danio rerio CV2 VWC1 domain in its unbound state showing that the key features for high affinity binding to BMP-2 is a pre-oriented peptide loop.},
  language  = {en}
}
@article{OnoSonoyamaNemaetal.2014,
  author    = {Ono, Mitsuaki and Sonoyama, Wataru and Nema, Kazuki and Hara, Emilio Satoshi and Oida, Yasutaka and Pham, Hai Thanh and Yamamoto, Katushi and Hirota, Kazuo and Sugama, Kazushige and Sebald, Walter and Kuboki, Takuo},
  title     = {Regeneration of calvarial defects with Escherichia coli-derived rhBMP-2 adsorbed in PLGA membrane},
  series = {Cells Tissues Organs},
  volume    = {198},
  journal   = {Cells Tissues Organs},
  number    = {5},
  issn      = {1422-6405},
  doi       = {10.1159/000356947},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-196680},
  pages     = {367 -- 376},
  year      = {2014},
  abstract  = {Objective: Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. Materials and Methods: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. Results: Release-profile testing showed that PLGA membrane could retain 94\% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2. Conclusion: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.},
  language  = {en}
}
@article{KlammertMuellerHellmannetal.2015,
  author    = {Klammert, Uwe and M{\"u}ller, Thomas D. and Hellmann, Tina V. and Wuerzler, Kristian K. and Kotzsch, Alexander and Schliermann, Anna and Schmitz, Werner and Kuebler, Alexander C. and Sebald, Walter and Nickel, Joachim},
  title     = {GDF-5 can act as a context-dependent BMP-2 antagonist},
  series = {BMC Biology},
  volume    = {13},
  journal   = {BMC Biology},
  number    = {77},
  doi       = {10.1186/s12915-015-0183-8},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-125550},
  year      = {2015},
  abstract  = {Background Bone morphogenetic protein (BMP)-2 and growth and differentiation factor (GDF)-5 are two related transforming growth factor (TGF)-β family members with important functions in embryonic development and tissue homeostasis. BMP-2 is best known for its osteoinductive properties whereas GDF-5—as evident from its alternative name, cartilage derived morphogenetic protein 1—plays an important role in the formation of cartilage. In spite of these differences both factors signal by binding to the same subset of BMP receptors, raising the question how these different functionalities are generated. The largest difference in receptor binding is observed in the interaction with the type I receptor BMPR-IA. GDF-5, in contrast to BMP-2, shows preferential binding to the isoform BMPR-IB, which is abrogated by a single amino acid (A57R) substitution. The resulting variant, GDF-5 R57A, represents a "BMP-2 mimic" with respect to BMP receptor binding. In this study we thus wanted to analyze whether the two growth factors can induce distinct signals via an identically composed receptor. Results Unexpectedly and dependent on the cellular context, GDF-5 R57A showed clear differences in its activity compared to BMP-2. In ATDC-5 cells, both ligands induced alkaline phosphatase (ALP) expression with similar potency. But in C2C12 cells, the BMP-2 mimic GDF-5 R57A (and also wild-type GDF-5) clearly antagonized BMP-2-mediated ALP expression, despite signaling in both cell lines occurring solely via BMPR-IA. The BMP-2- antagonizing properties of GDF-5 and GDF-5 R57A could also be observed in vivo when implanting BMP-2 and either one of the two GDF-5 ligands simultaneously at heterotopic sites. Conclusions Although comparison of the crystal structures of the GDF-5 R57A:BMPR-IAEC- and BMP-2:BMPR-IAEC complex revealed small ligand-specific differences, these cannot account for the different signaling characteristics because the complexes seem identical in both differently reacting cell lines. We thus predict an additional component, most likely a not yet identified GDF-5-specific co-receptor, which alters the output of the signaling complexes. Hence the presence or absence of this component then switches GDF-5′s signaling capabilities to act either similar to BMP-2 or as a BMP-2 antagonist. These findings might shed new light on the role of GDF-5, e.g., in cartilage maintenance and/or limb development in that it might act as an inhibitor of signaling events initiated by other BMPs.},
  language  = {en}
}
@article{SeherLaglerStuehmeretal.2017,
  author    = {Seher, Axel and Lagler, Charlotte and St{\"u}hmer, Thorsten and M{\"u}ller-Richter, Urs Dietmar Achim and K{\"u}bler, Alexander Christian and Sebald, Walter and M{\"u}ller, Thomas Dieter and Nickel, Joachim},
  title     = {Utilizing BMP-2 muteins for treatment of multiple myeloma},
  series = {PLoS ONE},
  volume    = {12},
  journal   = {PLoS ONE},
  number    = {5},
  doi       = {10.1371/journal.pone.0174884},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-158144},
  pages     = {e0174884},
  year      = {2017},
  abstract  = {Multiple myeloma (MM) represents a haematological cancer characterized by the pathological hyper proliferation of antibody-producing B-lymphocytes. Patients typically suffer from kidney malfunction and skeletal disorders. In the context of MM, the transforming growth factor β (TGFβ) member Activin A was recently identified as a promoter of both accompanying symptoms. Because studies have shown that bone morphogenetic protein (BMP)-2-mediated activities are counteracted by Activin A, we analysed whether BMP2, which also binds to the Activin A receptors ActRII and ActRIIB but activates the alternative SMAD-1/5/8 pathway, can be used to antagonize Activin A activities, such as in the context of MM. Therefore three BMP2 derivatives were generated with modified binding activities for the type II (ActRIIB) and/or type I receptor (BMPRIA) showing either increased or decreased BMP2 activity. In the context of MM these BMP2 muteins show two functionalities since they act as a) an anti-proliferative/apoptotic agent against neoplastic B-cells, b) as a bone-formation promoting growth factor. The molecular basis of both activities was shown in two different cellular models to clearly rely on the properties of the investigated BMP2 muteins to compete for the binding of Activin A to the Activin type II receptors. The experimental outcome suggests new therapeutic strategies using BMP2 variants in the treatment of MM-related pathologies.},
  language  = {en}
}