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. Author manuscript; available in PMC: 2012 Feb 18.
Published in final edited form as: Traffic. 2003 Mar;4(3):153–161. doi: 10.1034/j.1600-0854.2003.00103.x

Figure 2. Golgin-84 binds directly and specifically to rab1 GTPase.

Figure 2

A. Detergent extracts of RLG were incubated with GST-rab1 (lanes 2 and 3), GST-rab2 (lanes 4 and 5), GST-rab6 (lanes 6 and 7), GST-rab11 (lanes 8 and 9) or GST alone (lane 10). The rabs were preloaded with either GTPγS (lanes 2, 4, 6 and 8) or GDP (lanes 3, 5, 7 and 9). The input Golgi (10%) is shown in lane 1. Bound proteins were eluted with GDP/EDTA and fractionated by SDS-PAGE followed by Western blotting for golgin-84 (upper panel) or GM130 (lower panel). B. Soluble, recombinant golgin-84 was incubated with GST-rab1 (lanes 2 and 3) or GST-rab11 (lanes 4 and 5), preloaded with either GTPγS (lanes 2 and 4) or GDP (lanes 3 and 5). Incubations were also carried out using the ‘GTP’ (rab1 Q67 L) and ‘GDP’ (rab1 S25 N) locked forms of rab1 (lanes 6 and 7). The input Golgi (10%) is shown in lane 1. Bound proteins were analyzed by SDS-PAGE followed by Western blotting for Golgin-84 (WB), or by Coomassie Brilliant Blue (CBB) staining to show the amount of rabs used.