Figure 1. Mechanism of isoaspartate formation and PIMT-catalyzed repair.

Under physiological conditions, deamidation of asparagine residues or dehydration of aspartic acid residues results in the formation of a metastable intermediate succinimide which spontaneously hydrolyzes to form a mixture of normal L-aspartyl and atypical L-isoaspartyl linkages. PIMT, using AdoMet as a methyl donor, selectively methylates the isoaspartyl α-carboxyl group to form a highly labile methyl ester. Spontaneous demethylation occurs within minutes to reform the original succinimide, with release of methanol as a by-product. This succinimide is now the starting point for further cycles of repair, resulting in near complete conversion of the isoaspartyl β-linkages to normal aspartyl α-linkages.