Different H12 orientations in PPARγ LBD are superimposed in the
PPARγ LBD, as shown by an ensemble of structural models of different
orientations of H12 (K474) conjugated to K457 (blue H12, Figure 2C), K275 (red H12, Figure 2D), K265 (orange H12, Figure 2E) and K301 (green H12, Figure 2F). Yellow, H3 residue region SVEAVQEITEY;
Cyan: BS3 conjugated lysine residues shown in stick. (A) relative peak area of
non-XL peptide SVEAVQEITEY. (B) relative peak area of K224-K232 XL peptide.
Results are expressed as mean +/− SEM. (C-F) relative peak area of
K474-K457, K474-K275, K474-K265 and K474-K301 XL peptide, respectively. All
results are expressed as mean +/− SEM. Statistical analysis was performed
by two-way ANOVA between indicated pairwise experiment (*** = p<0.001; **
= p<0.01; * = p<0.05). DynaXL program is used to generate input
structure calculation scripts that is further used by the xplor-nih program for
structure modeling of XL-MS data. The data is consistent with models for both
PPARγ alone and the PPARγRXRα heterodimer complex. See also
Figure S3 and Figure
S4.