Reconstitution of Rab- And SNARE-dependent membrane fusion by synthetic endosomes
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
Abstract
Rab GTPases and SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are evolutionary conserved essential components of the eukaryotic intracellular transport system. Although pairing of cognate SNAREs is sufficient to fuse membranes in vitro, a complete reconstitution of the Rab-SN ARE machinery has never been achieved. Here we report the reconstitution of the early endosomal canine RabS GTPase, its key regulators and effectors together with SNAREs into proteoliposomes using a set of 17 recombinant human proteins. These vesicles behave like minimal 'synthetic' endosomes, fusing with purified early endosomes or with each other in vitro. Membrane fusion measured by content-mixing and morphological assays requires the cooperativity between RabS effectors and cognate SNAREs which, together, form a more efficient 'core machinery' than SNAREs alone. In reconstituting a fusion mechanism dependent on both a Rab GTPase and SNAREs, our work shows that the two machineries act coordinately to increase the specificity and efficiency of the membrane tethering and fusion process.
Details
Originalsprache | Englisch |
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Seiten (von - bis) | 1091-1097 |
Seitenumfang | 7 |
Fachzeitschrift | Nature |
Jahrgang | 459 |
Publikationsstatus | Veröffentlicht - 25 Juni 2009 |
Peer-Review-Status | Ja |
Extern publiziert | Ja |
Externe IDs
WOS | 000267636700035 |
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Scopus | 67649470529 |
PubMed | 19458617 |