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SmgGDS preferentially forms a high affinity complex with nucleotide-depleted RhoA possessing a polybasic region. SmgGDS was incubated on ice for 30 min with a 2-fold molar excess of RhoA loaded with GDP (A), GTPγS (B), or in the presence of EDTA (C), separated over size-exclusion chromatography, and fractions were analyzed via SDS-PAGE.
