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. Author manuscript; available in PMC: 2008 Sep 15.
Published in final edited form as: Biochemistry. 2007 Jan 9;46(1):271–278. doi: 10.1021/bi061858p

FIGURE 5.

FIGURE 5

Ime2 can phosphorylate proteins other than Sum1 that contain the RPST motif. Mbp fused to residues 1-211 of Gip1, or residues 1162-1447 of Sgs1, or the same proteins containing alanine (A) in place of the threonine (T) in the RPST motif as indicated, were incubated with Ime2 in the presence of radiolabeled ATP. (A) Autoradiograph of the reactions analyzed by electrophoresis. The migration of Mbp, Mbp-Gip1-1-211, Mbp-Sgs1-1162-1447 and of the Ime2 autophosphorylation product are indicated. (B) Coomassiestained gel of the protein substrates analyzed in parallel. Analysis of the unstable Mbp-Gip1-1-211 fusion protein demonstrates that all of the observable Coomassie-stainable protein bands contain Mbp as assayed by Western immunoblot analysis (data not shown).