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Updated: Jul 5, 2026

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
Published on: December 14, 2017
Identification and verification of Sro7p as an effector of the Sec4p Rab GTPase
Bianka L Grosshans1, Peter Novick
1Novartis Institutes for Biomedical Research, Basel, Switzerland.
Abstract:
Effectors are operationally defined as proteins that recognize a specific GTPase preferentially in its GTP-bound conformation. Here we present the use of affinity chromatography to identify potential effectors of Sec4p, the Rab GTPase that controls the final stage of the yeast secretory pathway. We describe the preparation of the Rab protein affinity matrix and the yeast lysate used in the purification. We also describe the methods used to identify and verify one candidate, Sro7p, as a bona fide Sec4p effector. This includes tests of the specificity and efficiency of binding both in vitro and in vivo.
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