The presence of neutral metalloproteolytic activity and metalloproteinase inhibitors in the interphotoreceptor matrix.
Neutral proteolytic activity, having a pH optimum of about 7, was present in the high molecular weight fraction of bovine interphotoreceptor matrix (IPM) separated by gel filtration on Sephacryl S-500. The enzyme(s) was active toward a number of exogenous substrates, including albumin, Azocoll, and gelatin. However, it was inactive toward a synthetic substrate for bacterial collagenase. Proteolytic activity was proportional to protein; however, the time course of the reaction was nonlinear, suggesting that "activation" of a precursor form might be necessary. Of a number of specific inhibitors tested, those directed toward metalloproteinases (1,10-phenanthroline greater than EDTA greater than EGTA) proved most effective. While activity was also inhibited by sulfhydryl reagents and dithiothreitol, inhibitors specific for cysteine proteinases were ineffective. Higher specific activity was present in IPM obtained from retinal pigment epithelium (RPE) than from retina. An endogenous proteinase inhibitor(s) was also found in IPM from both RPE and retina. It was effective against the endogenous metalloproteolytic activity of IPM and also against thermolysin, but not against trypsin or papain. Fractionation of IPM on Sephacryl S-500 revealed a broad peak of inhibitory activity at molecular weights of less than 10(5) daltons. This is the first report of the presence of neutral proteolytic activity and metalloproteinase inhibitor(s) in bovine IPM. These materials may function in concert to maintain the proper level of various components within this matrix.
Plantner
J J
JJ
Lorand V. Johnson Laboratory for Research in Ophthalmology, Department of Ophthalmology, Case Western Reserve University, Cleveland, OH 44106.
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000201:enzymology*; D010857:Pigment Epithelium of Eye / Q000201:enzymology; D013379:Substrate Specificity
D008666:Metalloendopeptidases