Figure 5.

SpCENP-A loading chaperone Sim3 requires GRANT prolines for binding. (A) Normalized emPAI scores of co-precipitated Sim3 by the Cnp1-4PA (4PA) peptide relative to wild type (WT) identified through mass spectrometric quantification. Numbers are the mean of two experiments (Normalized emPAI of 4PA = 0.54). (B) Sim3-GFP physically interacts with the SpCENP-A NTD peptide but the interaction is reduced for Cnp1–4PA. Cbh1-HA is used as a positive control and loading control. Representative blot of three experimental replicates. Graph depicts quantitative band intensities. Relative fold intensities are obtained via normalization with WT. Two tailed t-test, Error: S.D., *P < 0.05. Pept PD, peptide pull down (C) Isothermal titration calorimetry (ITC) profiles between Sim3-His and WT (left) or Cnp1-4PA peptides (right).