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. 2013 Sep 26;288(45):32466–32474. doi: 10.1074/jbc.M113.498329

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Kinetic analysis of the Rab8-GRAB interaction. A, comparison of full-length GRAB (GRAB_fl) GEF activity with the GRAB and Rabin8 GEF domains (10 μm) interacting with Rab8·mant-GDP (mGDP) in the presence of 100 μm GDP. Rab3·mant-GDP did not interact with full-length GRAB (10 μm). rel., relative; a. U., arbitrary units. B, dependence of the observed rate constant of mant-GDP/mant-GppNHp release from Rab8 on the concentration of GRAB. Plots of the observed rate constants for mant-GDP (black squares)/mant-GppNHp (gray squares) were hyperbolic. Fitting the hyperbolic curve leads to the values for K_D₃ and k″₋₄. C, dependence of the observed rate constants for the association reaction on the concentration of nucleotide-free Rab8·GRAB complex.