Polarized distribution of metalloproteinases in the bovine interphotoreceptor matrix.
Previous studies from this laboratory had indicated that metalloproteolytic activity was present in the interphotoreceptor matrix. This report extends those observations by providing evidence for the presence of multiple forms of metalloproteinase and for their polarized distribution within the interphotoreceptor matrix as determined by zymogram analysis on substrate-loaded gels. Retinal pigment epithelium-associated interphotoreceptor matrix, both unfractionated as well as fractions separated by gel filtration, exhibited bands of proteolytic activity on gelatin-loaded gels at about 70-75 kDa and 90 kDa, possibly due to gelatinases A and B (MMP-2 and MMP-9, respectively). In contrast, no neutral proteolytic activity was seen in retina-associated interphotoreceptor matrix unless it was first fractionated by gel filtration, whereupon a diversity of forms was exhibited, including additional major bands of proteolytic activity at about 150 kDa and 100 kDa, on gelatin-loaded gels, and at about 180 kDa, on casein-loaded gels, along with many minor species. In all cases, all proteinase activity was inhibited by chelating agents. Since these enzymes may be involved in the turnover and remodeling of components present in the interphotoreceptor matrix, many of which are distributed in a compartmentalized fashion, this unequal distribution of metalloproteinases may be correlated with substrate specificity.
Plantner
J J
JJ
Department of Ophthalmology, Case Western Reserve University, Cleveland, OH 44106, USA.
Drew
T A
TA
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000201:enzymology*; D018093:Gelatinases / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism*; D008970:Molecular Weight; D010786:Photoreceptor Cells / Q000201:enzymology*; D010857:Pigment Epithelium of Eye / Q000201:enzymology
D018093:Gelatinases; D008666:Metalloendopeptidases; C030895:caseinase