Per residue characteristics.

<p>(A) Residue root mean square fluctuations (RMSF) are provided for the six molecular dynamics trajectories. The increased fluctuations within the double and quadruple mutant trivially reflect the VH/VL global rearrangements at the dimer interface. Note that the linker region, indicated by the short tick marks along the x-axis, is ignored because the fluctuations therein obfuscate the rest. The dashed vertical lines indicate where mutations occur. (B) The average flexibility index is provided for each case. Reported values correspond to the appropriate weighted average (defined in methods) over 10 representative structures sampled from the MD trajectory. Changes in flexibility relative to the wild type are both structurally local (as observed at the two highlighted mutant locations) and remote from the mutation site. (C) Differences in H-bond between wild type and each mutant. The total H-bond counts (donor and acceptor) are averaged across the MD simulation for each residue. Note that there is no wild type (black) series in panel (C) because the reported values are differences.