Characterization of soluble guanylyl cyclase in transformed human non-pigmented epithelial cells.
Topical application of nitro vasodilators, such as sodium nitroprusside, reduces intraocular pressure. In brain and blood vessels, nitro vasodilators activate soluble guanylyl cyclases, producing cGMP. The location and molecular identity of ocular guanylyl cyclases are unknown. We studied transformed human non-pigmented ciliary epithelial cells, whose parental cells are responsible for the production of aqueous humor. Sodium nitroprusside increased cGMP 40 to 60 fold in a time and concentration dependent manner (EC50 40 to 100 microM). Methylene blue inhibited this effect (IC50 0.6 microM in the presence of 100 microM sodium nitroprusside). We also detected mRNA for the alpha 1 and beta 1, but not the beta 2, subunit isoforms of soluble guanylyl cyclase in these cells by Northern blotting. We conclude that transformed non-pigmented epithelial cells contain an active heterodimeric soluble guanylyl cyclase composed of at least alpha 1 and beta 1 subunits.
Danziger
R S
RS
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas.
Star
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Matsumoto
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Coca-Prados
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DeSantis
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Pang
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D015152:Blotting, Northern; D002461:Cell Line, Transformed; D006152:Cyclic GMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology; D006162:Guanylate Cyclase / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008751:Methylene Blue / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism
D007527:Isoenzymes; D046911:Macromolecular Substances; D012333:RNA, Messenger; D009599:Nitroprusside; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D008751:Methylene Blue