Measuring Rab GTPase-Activating Protein (GAP) Activity in Live Cells and Extracts
Mammalian cells encode a diverse set of Rab GTPases and their corresponding regulators. In vitro biochemical screening has proven invaluable in assigning particular Rabs as substrates for their cognate GTPase-activating proteins. However, in vitro activity does not always reflect substrate specificity in cells. This method describes a functional test of GAP activity in cells or extracts that takes into account the presence of other factors or conditions that might change observed in vitro specificity.
Key wordsGTPase-activating protein Rab GTPase Effector protein
This research was supported by a grant from the National Institutes of Health (NIDDK 37332-28) to SRP.