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. Author manuscript; available in PMC: 2012 Jul 1.
Published in final edited form as: Nat Struct Mol Biol. 2011 Dec 4;19(1):48–55. doi: 10.1038/nsmb.2178

Figure 6. Interaction of Ndc10 Domain IV–V with N-terminal Scm3.

Figure 6

(a) Ni2+ affinity pull-down of 35S-labeled, in vitro translated Ndc10 proteins with purified, His-tagged Scm3 proteins, analyzed by SDS-PAGE and visualized by phosphoimaging. (b) In vitro amylose pull down of purified MBP tagged Scm3 proteins with Ndc10 domain IV–V. (c) Schematic overview of domain association of K. lactis Ndc10 with other kinetochore proteins. Ndc10 DI interacts with CBF3 core; Ndc10 DI–II, with Cbf1 (229–359) and Bir1p (1–328). Scm3 (1–28) associates with Ndc10 DIV–V but not with DV. Interaction of Cbf1 with Ndc10 was confirmed by analytical size-exclusion chromatography with purified proteins (Supplementary Fig. 6).