Purification of active HOPS complex reveals its affinities for phosphoinositides and the SNARE Vam7p

Coupling of Rab GTPase activation and SNARE complex assembly during membrane fusion is poorly understood. The homotypic fusion and vacuole protein sorting (HOPS) complex links these two processes: it is an effector for the vacuolar Rab GTPase Ypt7p and is required for vacuolar SNARE complex assembly...

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Bibliographic Details
Published in:The EMBO journal 2006-04, Vol.25 (8), p.1579-1589
Main Authors: Stroupe, C, Collins, K.M, Fratti, R.A, Wickner, W
Format: Article
Language:English
Subjects:
binding capacity
binding proteins
Binding sites
Enzymes
fungal proteins
homotypic fusion and vacuole protein sorting complex
HOPS
Membrane Fusion
Molecular biology
phosphatidylinositols
Phosphatidylinositols - metabolism
phosphoinositide
phosphoinositides
Protein Binding
Protein Structure, Tertiary
protein-protein interactions
Proteins
Qc-SNARE Proteins - isolation & purification
Qc-SNARE Proteins - metabolism
Rab
rab GTP-Binding Proteins - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins - isolation & purification
Saccharomyces cerevisiae Proteins - metabolism
SNARE
SNARE protein
Synaptosomal-Associated Protein 25
vacuoles
Vacuoles - physiology
Vam7p protein
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Summary:Coupling of Rab GTPase activation and SNARE complex assembly during membrane fusion is poorly understood. The homotypic fusion and vacuole protein sorting (HOPS) complex links these two processes: it is an effector for the vacuolar Rab GTPase Ypt7p and is required for vacuolar SNARE complex assembly. We now report that pure, active HOPS complex binds phosphoinositides and the PX domain of the vacuolar SNARE protein Vam7p. These binding interactions support HOPS complex association with the vacuole and explain its enrichment at the same microdomains on docked vacuoles as phosphoinositides, Ypt7p, Vam7p, and the other SNARE proteins. Concentration of the HOPS complex at these microdomains may be a key factor for coupling Rab GTPase activation to SNARE complex assembly.
ISSN:0261-4189
1460-2075
DOI:10.1038/sj.emboj.7601051