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. Author manuscript; available in PMC: 2014 Nov 24.
Published in final edited form as: Nat Struct Mol Biol. 2013 Nov 3;20(12):1407–1414. doi: 10.1038/nsmb.2707

Figure 2.

Figure 2

Sequence requirements for Cks1 binding. (a) Dot-blot peptide experiment to determine the minimum length required for association with Cks1. The indicated peptides were conjugated to BSA, spotted onto PVDF membrane, incubated with His-tagged Cks1, and probed with an anti-His antibody. The data demonstrate the requirement of a 7-mer containing a phosphothreonine. The full blot for this experiment can be seen in Supplemental Figure 7a. (b) Scanning mutagenesis SPOT peptide array based on the Cdc63–10 AIPIpTPTK peptide. Peptides containing the indicated single-amino acid substitution were synthesized onto the membrane and probed for Cks1 binding as in (a). (c) Similar to (b), except the array is constructed from a phosphorylated Sic12–9 peptide.