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. Author manuscript; available in PMC: 2011 Sep 8.
Published in final edited form as: Curr Biol. 2011 Feb 25;21(5):391–398. doi: 10.1016/j.cub.2010.12.039

Figure 3. Wild type but not mutant Cenp-C1–71 binds Mis12C.

Figure 3

(A) Size-exclusion chromatography elution profiles and SDS-PAGE analysis of recombinant Mis12CMini (upper panel), recombinant GST-Cenp-C1–71 (middle panel), and their stoichiometric combination (lower panel). Complex formation is indicated by a shift in the elution profile of GST-Cenp-C1–71 and Mis12CMini and their appearance, in stoichiometric amounts, in early elution volumes. (B) Multiple sequence alignment of the N-terminal region of Cenp-CV from the indicated species. The colouring scheme reports the degree of conservation. The position of three predicted helices is shown on the sequence of human Cenp-C. (C) As in A, but with a GST-Cenp-C1–71Y13A mutant. The upper panel is the same as in A. No shift in elution volume is observed, indicating that binding is impaired. (D) As in A, but with a GST-Cenp-C1–71K10A mutant. The upper panel is the same as in A. Also with this mutant no shift in elution volume is observed.