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JoVE Journal
Biology

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Comparing the Affinity of GTPase-binding Proteins using Competition Assays
 

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Article DOI: 10.3791/53254
October 8th, 2015

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Summary October 8th, 2015

This protocol compares the relative affinities of binding partners for Rho-family GTPases, including Rac1. In vivo, Rac1-binding proteins compete for a single binding interface, the conformation of which is dictated by a bound nucleotide. The nucleotide is both important and difficult to control experimentally, due to the high hydrolysis rate.

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