論文 - 詳細
RRC ID | 28673 |
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著者 | Sato K, Nakano A. |
タイトル | Dissection of COPII subunit-cargo assembly and disassembly kinetics during Sar1p-GTP hydrolysis. |
ジャーナル | Nat Struct Mol Biol |
Abstract |
COPII coat proteins are required for direct capture of cargo and SNARE proteins into transport vesicles from the endoplasmic reticulum (ER). Cargo and SNARE capture occurs during the formation of a 'prebudding complex' comprising a cargo, Sar1p-GTP and the COPII subunits Sec23/24p. The assembly and disassembly cycle of the prebudding complex on ER membranes is coupled to the Sar1p GTPase cycle. Using FRET to monitor a single round of Sec23/24p binding and dissociation from SNAREs in reconstituted liposomes, we show that Sec23/24p dissociates from v-SNARE and complexed t-SNARE with kinetics slower than Sar1p-GTP hydrolysis. Once Sec23/24p becomes associated with v-SNARE or complexed t-SNARE, the complex remains assembled during multiple rounds of Sar1p-GTP hydrolysis mediated by the GDP-GTP exchange factor Sec12p. These data suggest a model for the maintenance of kinetically stable prebudding complexes during the Sar1p GTPase cycle that regulates cargo sorting into transport vesicles. |
巻・号 | 12(2) |
ページ | 167-74 |
公開日 | 2005-2-1 |
DOI | 10.1038/nsmb893 |
PII | nsmb893 |
PMID | 15665868 |
MeSH | COP-Coated Vesicles / metabolism* Fluorescence Resonance Energy Transfer GTP Phosphohydrolases / metabolism GTPase-Activating Proteins Guanine Nucleotide Exchange Factors Guanosine Diphosphate / metabolism Guanosine Triphosphate / metabolism* Hydrolysis Kinetics Liposomes / metabolism Membrane Glycoproteins / metabolism Membrane Proteins / metabolism Monomeric GTP-Binding Proteins / metabolism* Protein Subunits / metabolism Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae Proteins / metabolism* Time Factors Vesicular Transport Proteins |
IF | 11.98 |
引用数 | 95 |
WOS 分野 | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY |
リソース情報 | |
遺伝子材料 | pKSY193 (RDB08689) pKSY197 (RDB08690) pKSY198 (RDB08691) pET21a-SED5 (RDB08692) pKSE165 (RDB08708) pKSE168 (RDB08709) pKSE174 (RDB08710) pKSE176 (RDB08711) pKSE188 (RDB08712) pKSE203 (RDB08713) |