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. 2017 Dec 27;6:e33442. doi: 10.7554/eLife.33442

Figure 1. The interaction of CENP-N with nucleosomes.

(A) Schematic of crucial CCAN and KMN subunits discussed in the text. The Knl1-Mis12-Ndc80 (KMN) complex is the main microtubule receptor at the kinetochore. Other interactions are discussed in the main text. The question mark indicates that the precise determinants for the recruitment of CENP-LN to CENP-C and for the interaction of CENP-N with the CENP-A nucleosome have not been identified. (B) Schematic depicting constructs described in the manuscript. (C–E) Solid phase binding assays where the indicated GST fusion proteins were immobilized on glutathione-sepharose beads (at a final concentration of 1 µM) and incubated with 3 µM of the indicated nucleosome core particles. After incubation (see Materials and methods), beads were centrifuged, washed, and bound proteins visualized by SDS-PAGE and Coomassie staining.

Figure 1.

Figure 1—figure supplement 1. Further interactions of CENP-NL.

Figure 1—figure supplement 1.

Solid phase binding assay demonstrating that CENP-LN230-C does not interact with the CENP-A nucleosome. The indicated GST fusion proteins were immobilized on glutathione-sepharose beads (at a final concentration of 1 µM) and incubated with 3 µM of the indicated nucleosome core particles. After incubation (see Materials and methods), beads were pelleted, washed, and bound proteins visualized by SDS-PAGE and Coomassie staining.