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. Author manuscript; available in PMC: 2010 Jul 14.
Published in final edited form as: Biochemistry. 2009 Jul 14;48(27):6390–6401. doi: 10.1021/bi900621w

Figure 7. Arg-1261 is not necessary for larger truncations of Vps15 to bind to Gpa1.

Figure 7

(A) Detergent-solubilized extracts (Total) from cells expressing the indicated Myc fusion proteins and Gpa1 or Gpa2 fused to GST were incubated with glutathione-Sepharose resin, eluted with glutathione (GST PD), resolved by 10% SDS-PAGE, and analyzed by immunoblotting with antibodies against Myc, GST, Ste4 and G6PDH (Load control). (B) Detergent-solubilized extracts from cells expressing the indicated Myc fusion proteins and Gpa1 fused to GST were lysed in the presence of either GDP or GDP-AlF4-, incubated with glutathione-Sepharose resin, eluted with glutathione, and analyzed by immunoblotting with antibodies against Myc, GST, Ste 4, and G6PDH. PD, pulldown. GST, glutathione S-transferase. RA, Arg-1261 to Ala. RK, Arg-1261 to Lys.