Age-related changes in calpain II and calpastatin in rat lens

The purpose of these experiments was to determine how the activity and regulation of calpain in rat lens changed during aging. Calpain Ii enzyme activity and immunoreactivity decreased with both chronological and anatomical age. Two pieces of data suggested that loss of soluble calpain II was a result of both autolysis and insolubilization during aging: (i) proteolytic fragments of calpain were detected in lenses with molecular weights similar to fragments produced during incubation of purified calpain II with calcium; (ii) the water-insoluble fraction of lens cortex contained increasing amounts of calpain antigen durign aging, both the 75-kDa calpain subunit and a unique high-molecular-weight immunoreactive protein. The regulation of calpain II also appeared to change with age. The activity of calpain II in vivo may be regulated by the relative concentrations of calpain II and its endogenous inhibitor calpastatin. Calpain II concentrations decreased in the rat lens with age, whereas levels of the endogenous inhibitor calpastatin were maintained. Assays of calpain II and calpastatin indicated that upon aging there was insufficient activity of calpain II to overcome the inhibition of calpastatin in the nucleus. These findings were confirmed by incubation of crude lens homogenates of 2-week- and 7-month-old rat lens regions with calcium. It is hypothesized that binding of calpain II to membrane may be important for calpain II activation, especially in older lens regions, because it may allow escape from the inhibitory action of calpastatin.