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. Author manuscript; available in PMC: 2010 May 29.
Published in final edited form as: Cell. 2009 May 29;137(5):938–948. doi: 10.1016/j.cell.2009.03.043

Figure 7. RhoBTB3 is a Rab9-regulated ATPase that binds TIP47.

Figure 7

(A). ATP hydrolysis was determined at the times indicated using 100nM active His-RhoBTB3 full length protein or GST-Rho domain, in the absence or presence of either Rab9 (7.6μM active protein) or Rab5 (8.25μM active protein) and 50μM ATP. Right inset, SDS-PAGE analysis of proteins used. (B). Immunoblot analysis of blue native gel electrophoresis of K562 Golgi/endosome-enriched membranes or K562 cytosol (10μg each). Native protein marker migration is shown at left (kD). Left panel, RhoBTB3; right panel, TIP47. X-link indicates samples that were reacted with DTSSP crosslinker. (C). Rab9 and ATP stimulate release of TIP47 from the RhoBTB3 complex. Top panel, representative immunoblot; below, quantitation of duplicate experiments, with standard deviation.