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. Author manuscript; available in PMC: 2013 Mar 14.
Published in final edited form as: Antioxid Redox Signal. 2007 Nov;9(11):1803–1813. doi: 10.1089/ars.2007.1579

FIG. 4. CA-MKK6 and WT-MKK6 enhance Rac-GTPase activity in vitro.

FIG. 4

(A) Autoradiography of a representative thin-layer chromatography plate (TLC) separating the free 32phosphate [32Pi] from the [32P]γ-labeled GTP after hydrolysis by Rac1. Rac1 GTPase assays were performed in the presence or absence of GST-tagged CA-MKK6. Rac1 was preloaded with [32P]γ-GTP, and aliquots of the reaction were analyzed at various time points by thin-layer chromatography for GTP hydrolysis by assessing the percentage [32Pi] liberated from [32P]γ-GTP by Rac1. (B) Quantification of the percentage of GTP hydrolysis by Rac1 over time in the presence or absence of GST-tagged p50Rho-GAP (p29GAP), CA-MKK6, or boiled CA-MKK6. The graphs shown are representative of three independent experiments. (C) Quantification of the rate of GTP hydrolysis by Rac1 in the presence or absence of GST or the indicated GST-tagged MKK6 mutants. Values depict the mean ± SEM (n = 3). *Significantly different from Rac1 alone group; †significantly different from CA-MKK6 and WT-MKK6 groups (for all comparisons, p < 0.01; Student’s t test).