TY  - JOUR
A1  - Zhan, Hong
A1  - Stanciauskas, Ramunas
A1  - Stigloher, Christian
A1  - Dizon, Kevin K.
A1  - Jospin, Maelle
A1  - Bessereau, Jean-Luis
A1  - Pinaud, Fabien
T1  - In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans
JF  - Nature Communications
N2  - Single-molecule (SM) fluorescence microscopy allows the imaging of biomolecules in cultured cells with a precision of a few nanometres but has yet to be implemented in living adult animals. Here we used split-GFP (green fluorescent protein) fusions and complementation-activated light microscopy (CALM) for subresolution imaging of individual membrane proteins in live Caenorhabditis elegans (C. elegans). In vivo tissue-specific SM tracking of transmembrane CD4 and voltage-dependent Ca(2+) channels (VDCC) was achieved with a precision of 30 nm within neuromuscular synapses and at the surface of muscle cells in normal and dystrophin-mutant worms. Through diffusion analyses, we reveal that dystrophin is involved in modulating the confinement of VDCC within sarcolemmal membrane nanodomains in response to varying tonus of C. elegans body-wall muscles. CALM expands the applications of SM imaging techniques beyond the petri dish and opens the possibility to explore the molecular basis of homeostatic and pathological cellular processes with subresolution precision, directly in live animals.
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-121125
VL  - 5
IS  - 4974
ER  - 
TY  - JOUR
A1  - Rozycka, Miroslawa
A1  - Wojtas, Magdalena
A1  - Jakob, Michal
A1  - Stigloher, Christian
A1  - Grzeszkowiak, Mikolaj
A1  - Mazur, Maciej
A1  - Ozyhar, Andrzej
T1  - Intrinsically Disordered and Pliable Starmaker-Like Protein from Medaka (Oryzias latipes) Controls the Formation of Calcium Carbonate Crystals
JF  - PLOS ONE
N2  - Fish otoliths, biominerals composed of calcium carbonate with a small amount of organic matrix, are involved in the functioning of the inner ear. Starmaker (Stm) from zebrafish (Danio rerio) was the first protein found to be capable of controlling the formation of otoliths. Recently, a gene was identified encoding the Starmaker-like (Stm-l) protein from medaka (Oryzias latipes), a putative homologue of Stm and human dentine sialophosphoprotein. Although there is no sequence similarity between Stm-l and Stm, Stm-l was suggested to be involved in the biomineralization of otoliths, as had been observed for Stm even before. The molecular properties and functioning of Stm-l as a putative regulatory protein in otolith formation have not been characterized yet. A comprehensive biochemical and biophysical analysis of recombinant Stm-l, along with in silico examinations, indicated that Stm-l exhibits properties of a coil-like intrinsically disordered protein. Stm-l possesses an elongated and pliable structure that is able to adopt a more ordered and rigid conformation under the influence of different factors. An in vitro assay of the biomineralization activity of Stm-l indicated that Stm-l affected the size, shape and number of calcium carbonate crystals. The functional significance of intrinsically disordered properties of Stm-l and the possible role of this protein in controlling the formation of calcium carbonate crystals is discussed.
KW  - circular-dichroism
KW  - unstructured proteins
KW  - olyelectrolyte domains
KW  - modulating
KW  - biominarlization proteins
KW  - nacreous layer formation
KW  - alpha-helical structure
KW  - dye stains-all
KW  - polyelectrolyte domains
KW  - phosphorylation sites
KW  - procambarus-clarkii
KW  - secondary structure
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-114251
SN  - 1932-6203
VL  - 9
IS  - 12
ER  -