Kinetic study of racemization of aspartyl residues in recombinant human αA-crystallin

Abstract

Asp58 and Asp151 in human lens αA-crystallin invert and isomerize to d-β-aspartyl residues with age. Here, we report that the racemization rate constants ( k) of Asp58 and Asp151 residues in human recombinant αA-crystallin at 37 °C are 3.72 ± 0.8 × 10 −4 and 10.7 ± 0.7 × 10 −4/day, respectively. The activation energy of racemization of Asp58 and Asp151 in the protein was 27.0 ± 0.5 kcal/mol and 21.0 ± 0.5 kcal/mol, respectively. The time required for the D/ L ratio of Asp58 and Asp151 to approximate to 1.0 ( D/ L ratio of Asp = 0.99) at 37 °C was estimated as 20.9 ± 3.7 and 6.80 ± 0.4 years, respectively. Thus, Asp151 is more susceptible to racemization than Asp58, consistent with data from short model peptides. However, the racemization rates of both Asp58 and Asp151 residues in the protein were twice as rapid as in model peptides. These results indicate that the racemization of Asp residues in αA-crystallin may be influenced not only by the primary structure but also by the higher order structure around Asp residues in the protein.