Assaying the Interaction of the Rab Guanine Nucleotide Exchange Protein Sec2 with the Upstream Rab, a Downstream Effector, and a Phosphoinositide
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.
Key wordsSec2p Sec4p Sec15p Ypt32p Phosphatidylinositol-4-phosphate Guanine nucleotide exchange factor Effector
This work was supported by grants GM35370 and GM82861 from the N.I.H. to P.N. D.S was supported by Swiss National Science Foundation Grants PBSKP3_138593 and PBSKP3_145803.
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