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. 2015 May 12;13(5):e1002146. doi: 10.1371/journal.pbio.1002146

Fig 4. Molecular analysis of Cbk1 docking peptides in Ace2 and Ssd1 highlights importance of a [YF]xFP motif.

Fig 4

(A) Protein schematics with points of interest highlighted. Orange lines denote Cbk1 consensus sites, and blue boxes denote docking motifs. (B) Pulldown of Cbk1 kinase domain by truncated Ssd1 N-terminal (1–5) and C-terminal (6–11) docking motifs suggests that only the FKFP motif is required for interaction. (C) Alanine scan of the Ace2270–290 docking motif suggests that residues N-terminal to the YQFP motif aid in Cbk1 binding. (D) Mutational analysis of Ssd1 N-terminal docking motif highlights the sequence stringency of the core motif and suggests a consensus docking motif of [YF][KR]FP. (E) Cbk1 in vitro kinase assay with Ace2 or Ssd1 fragments containing a phosphorylation site (HxRxx[ST]) and either a WT docking motif (left) or mutated docking motif (dock*, right). Phosphorylation is enhanced in the presence of the WT docking motif.