Misalignment of the hGOT1 dimer interface determined by the first principal component (PC1).

A) Visualization of PC1 positive displacement (shown by yellow arrows) from PCA. Negative displacements correspond to the opposite direction of the arrows. The protein representation is the average coordinates of the inner Cα atoms (see Methods). B) The PC1 displacement of each variant as a function of simulation time. This displacement represents the magnitude of dimer interface misalignment. C) The monomer energy (EM) as a function of PC1 displacement. Ellipses show the standard deviation on both axes, and the mean values are located at the centers. D) and E) show the dimerization (EE) and PLP binding (EL) free energy versus PC1 displacement, respectively. The variants and WT follow the same color scheme as Fig 1. All values are relative to WT.