Loss of Function. The active site residues of PGM1 relatives have been annotated in the CDD database based on the 3D protein structure for PGM from Paramecium tetraurelia. In PAINT, the biocurator used the integrated multiple sequence alignment viewer to determine that key active site residues are mutated in all of the vertebrate PGM5 orthologs, suggesting that phosphoglucomutase activity was lost shortly after duplication. The biocurator correspondingly annotated the vertebrate ancestor of PGM5 with ‘NOT phosphoglucomutase activity’, which PAINT then propagated to all vertebrate orthologs of PGM5.