@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{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{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{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{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{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{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{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{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{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{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{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{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{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{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{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{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{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{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{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} }