Reconstitution of Rab- and SNARE-dependent membrane fusion by synthetic endosomes.
Nature
; 459(7250): 1091-7, 2009 Jun 25.
Article
em En
| MEDLINE
| ID: mdl-19458617
ABSTRACT
Rab GTPases and SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are evolutionarily 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-SNARE machinery has never been achieved. Here we report the reconstitution of the early endosomal canine Rab5 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 Rab5 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.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Endossomos
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Proteínas rab de Ligação ao GTP
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Proteínas SNARE
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Fusão de Membrana
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article