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. 2021 Jan 19;40(5):e105671. doi: 10.15252/embj.2020105671

Figure EV2. Cryo‐EM structure of the CA‐CCCT complex.

Figure EV2

  1. Cryo‐EM density map of the CENPC‐CT bound to the CENP‐A nucleosome at a 6.78 Å resolution. The EM map for CENPC‐CT is shown in a pink surface representation together with the ribbon representation of the CENP‐A nucleosome. The molecules in the complex are color‐coded as indicated in the figure. The right panel shows a slice view along the two‐fold axis. The two CENPC‐CT fragments symmetrically bind to the CENP‐A nucleosome.
  2. Comparison of lower (6.78 Å) and higher (4.5 Å) resolution EM densities of the CA‐CCCT complex. The map at 4.5 Å resolution is depicted in a surface representation and superposed on the 6.78 Å resolution map shown as a mesh representation (light gray). The superposed maps corresponding to the CENPC‐CT are enlarged in a left panel.
  3. Detailed views of the cryo‐EM density map of the CA‐CCCT complex at a 4.5 Å resolution. The map is shown as a mesh representation with the ribbon model of the final cryo‐EM structure.
  4. Protease sensitivity of CENPC‐CT is altered by the presence of the CENP‐A nucleosome. Schematic diagram showing Factor Xa cleavage sites in MBP‐fused chicken CENPC‐CT with its functional domain organization. The possible minor cleavage sites (Gly‐Arg sequence) are indicated as cutting site 2 (between residues 642 and 643) and 3 (between residues 682 and 683), in addition to a major cleavage site between CENPC‐CT and MBP (site 1). The amino acid sequence of the folded region in CENPC‐CT is indicated below. The lower left panel shows the result of SDS–PAGE analysis of limited proteolysis product of CENPC‐CT in the absence or presence of the CENP‐A nucleosome. Possible fragments generated by Factor Xa digestion are shown in the lower right panel. Bands corresponding to each fragment are indicated in the gel. In the absence of the CENP‐A nucleosome, bands of the limited proteolysis products (b, c, e, f, and g) were observed. These bands were not observed in the presence of CENP‐A nucleosome.