Peptide hydrolysis in lens: role of leucine aminopeptidase, aminopeptidase III, prolyloligopeptidase and acylpeptidehydrolase.

The distribution of leucine aminopeptidase, aminopeptidase III, prolyloligopeptidase and acylpeptidehydrolase activities in different regions of a bovine lens was determined and correlated with the distribution of crystallin fragments (measured as < 18 kDa protein) and water-insoluble proteins in the same lens. A gradient of activity was observed for all the peptidases tested, with the highest specific activity present in the cortical fibers which decreased to one half or below in the inner cortical fibers and nucleus. An inverse correlation between peptidase activities and the amount of crystallin fragments was observed in different regions of the lens. However, a direct correlation between the water-insoluble protein content and the crystallin fragments was observed in all fibers of the same lens. The amount of crystallin fragments and the amount of water-insoluble proteins increased from 2.7% and 8% in the outer cortical fibers to 13% and 68% in the nucleus of the same lens. The water-insoluble fraction from both cortical and nuclear fibers however displayed 4-5 fold more crystallin fragments compared to that present in the water-soluble fraction of the same preparation. When the bovine lens cortical and nuclear extracts were tested for their ability to hydrolyze the peptide substrate, Ile-Ser-bradykinin, the cortical extract was found to be at least ten times superior to the nuclear extract. Prior inactivation of prolyloligopeptidase and other serine proteases by diisopropylfluorophosphate however diminished the ability of the cortical extract to hydrolyze peptide substrates. Bovine lens cortical extract was able to completely hydrolyze alpha-melanocyte stimulating hormone as well as N-Acetyl-Met-Asp-Arg-Val-Leu-Ser-Arg-Tyr showing the presence of active acylpeptidehydrolase facilitating the complete hydrolysis of N-terminally blocked peptides. The human lens extract was found to contain both diisopropylfluorophosphate sensitive and resistant enzymes capable of hydrolyzing peptide substrates.