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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Nature. 2017 Feb 1;542(7640):255–259. doi: 10.1038/nature21061

Figure 4.

Figure 4

Biochemical characterizations of the interactions between separase and securin. (a). Domain organizations of S. cerevisiae and Z. rouxii securin. The KEN motif and the destruction box (D-box) are also indicated. (b). Z. rouxii separase (with an N-terminal His tag) was co-expressed with various segments of Z. rouxii securin. The eluates from nickel columns were separated by SDS gel electrophoresis. The positions of separase and securin are indicated with the black and red arrowheads, respectively. WT: full-length Z. rouxii separase; WT-CS: full-length Z. rouxii separase C1497S mutant; WT′: Z. rouxii separase with an internal deletion of residues 952–1010, corresponding to a poorly conserved, disordered loop in domain IV of the S. cerevisiae separase structure. For gel source data, see Supplementary Figure 1. (c). Summary of the expression results in panel b. The solubility levels of separase for various securin segments are indicated.