Characterization and novel activation of 72-kDa metalloproteinase in retinal interphotoreceptor matrix and Y-79 cell culture medium

Jones, B. Eric, Moshyedi, Payman, Gallo, Samuel, Tombran-Tink, Joyce, Arand, Gloria, Reid, Deborah A., Thompson, Erik W., Chader, Gerald J., & Waldbillig, Robert J. (1994) Characterization and novel activation of 72-kDa metalloproteinase in retinal interphotoreceptor matrix and Y-79 cell culture medium. Experimental Eye Research, 59(3), pp. 257-269.

View at publisher

Abstract

Analysis of bovine interphotoreceptor matrix and conditioned medium from human Y-79 retinoblastoma cells by gelatin SDS-PAGE zymography reveals abundant activity of a 72-kDa M(r) gelatinase. The 72-kDa gelatinase from either source is inhibited by EDTA but not aprotinin or NEM, indicating that it is a metalloproteinase (MMP). The 72-kDa MMP is converted to a 62-kDa species with APMA treatment after gelatin sepharose affinity purification typical of previously described gelatinase MMP-2. The latent 72-kDa gelatinase from either bovine IPM or Y-79 media autoactivates without APMA in the presence of calcium and zinc after 72 hr at 37°C, producing a fully active mixture of proteinase species, 50 (48 in Y-79 medium), 38 and 35 kDa in size. The presence of inhibitory activity was examined in both whole bovine IPM and IPM fractions separated by SDS-PAGE. Whole IPM inhibited gelatinolytic activity of autoactivated Y-79-derived MMP in a dose-dependent manner. Inhibitory activities are observed in two protein fractions of 27-42 and 20-25 kDa. Western blots using antibodies to human tissue inhibitor of metalloproteinase 1 and 2 (TIMP-1 and -2) reveal the presence of two TIMP-1-like proteins at 21 and 29 kDa in inhibitory fractions of the bovine IPM. TIMP-2 was not detected in the inhibitory IPM fractions, consistent with the observed autoactivation of bovine IPM 72-kDa gelatinase. Potential roles for this IPM MMP-TIMP system include physiologic remodelling of the neural retina-RPE cell interface and digestion of shed rod outer segment as well as pathological processes such as retinal detachment, PE cell migration, neovascularization and tumor progression. Cultured Y-79 cells appear to be a good model for studying the production and regulation of this proteinase system.

Impact and interest:

16 citations in Scopus
Search Google Scholar™
18 citations in Web of Science®

Citation counts are sourced monthly from Scopus and Web of Science® citation databases.

These databases contain citations from different subsets of available publications and different time periods and thus the citation count from each is usually different. Some works are not in either database and no count is displayed. Scopus includes citations from articles published in 1996 onwards, and Web of Science® generally from 1980 onwards.

Citations counts from the Google Scholar™ indexing service can be viewed at the linked Google Scholar™ search.

ID Code: 72179
Item Type: Journal Article
Refereed: Yes
Additional Information: Cited By (since 1996):15
Export Date: 6 May 2014
Source: Scopus
PubMed ID: 7821370
Additional URLs:
Keywords: Gelatinase, Inter-photoreceptor matrix, Metalloproteinase, Stromelysin, Tissue inhibitor of metalloproteinase, Y-79 retinoblastoma cell
DOI: 10.1006/exer.1994.1107
ISSN: 0014-4835
Divisions: Current > QUT Faculties and Divisions > Faculty of Health
Deposited On: 28 May 2014 01:54
Last Modified: 28 May 2014 01:54

Export: EndNote | Dublin Core | BibTeX

Repository Staff Only: item control page