Validation of residue conservation
Specific residues of CENP-H and CENP-M have been reported in different studies as essential in facilitating intermolecular interaction with residues of other subunits in the complex [14, 37]. We validated the evolutionary conservation of these residues with the use of ConSurf [38]. Consurf is popularly used for the detection of macromolecules’ functional regions, through the analysis of the evolutionary dynamics of nucleic acid and amino acid substitutions in homologous sequences. The tool evaluates the nucleic acid and amino acid evolutionary rates by mapping them onto the structure or sequence of the query macromolecule. Slowly evolving regions on the surface of the query macromolecule are known to be essential for functionality and thus, the analysis of ConSurf can highlight very important regions within the query macromolecule [38].