Abstract
Rabs are activated by guanine nucleotide exchange proteins, which are in turn controlled by complex regulatory mechanisms. Here we describe several different assays that have been used to delineate the mechanisms by which Sec2p, the exchange factor for the Rab Sec4p, is regulated. These assays assess the interaction of Sec2p with the upstream Rab, Ypt32p, a downstream Sec4p effector, Sec15p, and the lipid, phosphatidylinositol-4-phosphate.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Guanine Nucleotide Exchange Factors / genetics
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Guanine Nucleotide Exchange Factors / isolation & purification
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Guanine Nucleotide Exchange Factors / metabolism
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Immunoprecipitation
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Phosphatidylinositol Phosphates / metabolism*
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Protein Binding
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Protein Interaction Mapping / methods*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / isolation & purification
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Saccharomyces cerevisiae Proteins / metabolism*
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Vesicular Transport Proteins / genetics
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Vesicular Transport Proteins / isolation & purification
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Vesicular Transport Proteins / metabolism
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rab GTP-Binding Proteins / genetics
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rab GTP-Binding Proteins / isolation & purification
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rab GTP-Binding Proteins / metabolism*
Substances
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Guanine Nucleotide Exchange Factors
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Phosphatidylinositol Phosphates
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SEC15 protein, S cerevisiae
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SEC2 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Vesicular Transport Proteins
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phosphatidylinositol 4-phosphate
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YPT32 protein, S cerevisiae
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rab GTP-Binding Proteins