biostudies-literatureJoe MKNEI NIH HHS5308-5318https://www.ebi.ac.uk/biostudies/studies/S-EPMC5644706biostudies-literatureUnknown58(12)<h4>Purpose</h4>To elucidate functions of wild-type myocilin, a secreted glycoprotein associated with glaucoma.<h4>Methods</h4>Lysates of mouse eyes were used for immunoprecipitation with affinity-purified antibodies against mouse myocilin. Shotgun proteomic analysis was used for the identification of proteins interacting with myocilin. Colocalization of myocilin and tissue inhibitor of metalloproteinases 3 (TIMP3) in different eye structures was investigated by a multiplex fluorescent in situ hybridization and immunofluorescent labeling with subsequent confocal microscopy. Matrix metalloproteinase 2 (MMP2) activity assay was used to test effects of myocilin on TIMP3 inhibitory action.<h4>Results</h4>TIMP3 was identified by a shotgun proteomic analysis as a protein that was coimmunoprecipiInvestigative ophthalmology & visual scienceMyocilin Regulates Metalloprotease 2 Activity Through Interaction With TIMP3.PMC5644706R01 EY021205Nakaya NLieberman RLTomarev SIJoe MKfalseMyocilin Regulates Metalloprotease 2 Activity Through Interaction With TIMP3.<h4>Purpose</h4>To elucidate functions of wild-type myocilin, a secreted glycoprotein associated with glaucoma.<h4>Methods</h4>Lysates of mouse eyes were used for immunoprecipitation with affinity-purified antibodies against mouse myocilin. Shotgun proteomic analysis was used for the identification of proteins interacting with myocilin. Colocalization of myocilin and tissue inhibitor of metalloproteinases 3 (TIMP3) in different eye structures was investigated by a multiplex fluorescent in situ hybridization and immunofluorescent labeling with subsequent confocal microscopy. Matrix metalloproteinase 2 (MMP2) activity assay was used to test effects of myocilin on TIMP3 inhibitory action.<h4>Results</h4>TIMP3 was identified by a shotgun proteomic analysis as a protein that was coimmunoprecipi2017-01-01T00:00:00Z2017 Oct2025-04-22T09:10:36.075Z2019-03-27T02:59:09ZS-EPMC56447062904972910.1167/iovs.16-20336