Figure 3.
H/D Exchange Mass Spectrometry and Protein Pull-Downs Identify Interaction Domains between POLE3-POLE4 and H3-H4 Complexes
(A) Difference plot of hydrogen/deuterium (H/D) exchange data from POLE3 (upper left), POLE4 (upper right), H3 (lower left), and H4 (lower right). A cartoon depicting the domains of the analyzed proteins is shown on the top of the graph. Experimental error is reported in gray, while different colors represent different D2O incubation times. More exposed and protected regions are located, respectively, on the higher and lower part of the graph. Peptide aa numbers from highly protected or exposed regions of POLE3 and H4 are shown. ±0.5 Dalton difference is indicated with red/blue lines, which represents 98% confidence limit, so that peptides above or below these lines can be considered significantly changed. Difference heatmaps are provided in Figure S3A.
(B) Analytical gel filtration of the described protein complexes in 300 mM NaCl concentrations.
(C) SDS-PAGE analysis of gel filtration fractions from the described protein complexes.