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. 2018 Feb 5;217(2):601–617. doi: 10.1083/jcb.201705068

Figure 5.

Figure 5.

TRAPPII complex activates the Rab GTPases Rab1 and Rab11, whereas TRAPPIII only shows GEF activity toward Rab1. (A) Coomassie blue–stained protein gels of recombinant Drosophila TRAPP complexes purified from Sf9 cells coexpressing the subunits of each complex. FLAG tags on C10 or C11 allowed isolation of TRAPPII or TRAPPIII, respectively. PreScission protease (GST-HRV-3C protease) was used to cleave the tags (asterisks) and was subsequently removed using glutathione Sepharose beads. C10 also copurified with C9 in the absence of the shared subunits (TRAPPII lane). The Hsc70 chaperone (CG4264) is a contaminant of the TRAPPII purification protocol. Molecular masses are given in kilodaltons. (B) Release of mant-GDP from 250 nM of Rab-His6 by 50 nM of TRAPPII or TRAPPIII in the presence or absence of synthetic fly Golgi mix liposomes. Traces are the mean of at least three experiments. Error bars show SEM.