Functional reconstitution of ypt7p GTPase and a purified vacuole SNARE complex.
Membrane trafficking has heretofore been studied with intact organelles. Here, fusion-competent proteoliposomes were reconstituted from a yeast vacuole detergent extract. Homotypic vacuole fusion requires many membrane proteins, including the Ypt7p guanosine triphosphatase and a "SNARE complex" with Vam3p and Nyv1p. Proteoliposomes from extracts immunodepleted of either Vam3p or Ypt7p could ... not fuse, but vesicles reconstituted from a mixture of these depleted extracts had restored fusion activity. Purified preassembled vacuolar SNARE complex, when reconstituted with a SNARE-depleted extract, was fully functional for fusion. Thus, solubilized integral membrane components can be reconstituted for priming, docking, and fusion steps of organelle trafficking.
Mesh Terms:
Adenosine Triphosphate, Fungal Proteins, GTP Phosphohydrolases, GTP-Binding Proteins, Membrane Fusion, Membrane Proteins, Proteolipids, Qa-SNARE Proteins, SNARE Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Solubility, Vacuoles, Vesicular Transport Proteins, rab GTP-Binding Proteins
Adenosine Triphosphate, Fungal Proteins, GTP Phosphohydrolases, GTP-Binding Proteins, Membrane Fusion, Membrane Proteins, Proteolipids, Qa-SNARE Proteins, SNARE Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Solubility, Vacuoles, Vesicular Transport Proteins, rab GTP-Binding Proteins
Science
Date: Jul. 31, 1998
PubMed ID: 9685264
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