Effect of acetylation by aspirin on the thermodynamic stability of lens crystallins.

To assess the effect of aspirin on cataractogenesis, we compared the stability of individual, native protein fractions alpha L, beta H, beta L, beta s, beta B2, gamma-II, gamma-III and gamma-IV with that of their acetylated counterparts. The conformational stabilities of native fractions beta B2 and beta s, which were not reported earlier, were determined first from their thermal and a thermal denaturation behaviour. Since alpha L, beta H and beta L fractions are oligomeric, no thermodynamic analysis of these fractions was attempted. The thermal stability of beta s and beta B2 is rather low; their melting temperature (T1/2) range is 58-60 degrees C compared with 67-75 degrees C for the gamma-crystallins. Furthermore, except for alpha L, which remains stable even at 100 degrees C, and beta B2, all crystallins aggregate at temperatures slightly above T1/2. The Gibbs free energy of unfolding, delta GH2OD, calculated from guanidine HCl (GdnHCl) denaturation, is surprising low (3-9 kcal mol-1) for all crystallin fractions. The low values of delta GH2OD indicate that the structural destabilization of these proteins, which may lead to cataract formation, could result from a slight disturbance of a particular kind (sugar, UV light, oxidation, and other factors). The overall effect of acetylation on the individual crystallin fractions is mixed. The thermal stability of beta B2 increased, tended to decrease in the case of gamma-crystallins, but remained virtually unchanged for other proteins. Delta GH2OD values of the native crystallin fractions do not differ significantly from those of their acetylated counterparts.