title: Characterization of a novel monoclonal antibody that senses nitric oxide-dependent activation of soluble guanylate cyclase.
abstract: Two monoclonal antibodies (mAbs) against bovine lung soluble guanylate cyclase (sGC) were prepared and characterized. mAb 3221 recognized both the alpha- and beta-subunits of sGC and had greater binding affinity to the enzyme in the presence of NO. mAb 28131 recognized only the beta-subunit and its affinity did not change with NO. Neither mAb cross-reacted with particulate GC. Cultured Purkinje cells from rats were treated with S-nitroso-N-acetylpenicillamine, an NO donor, and examined by immunocytochemical methods. The immunoreactivity associated with mAb 3221 increased with the cGMP content in a crude extract of cerebellum and the NO2 generated in the culture medium increased.
authors.affiliation: Department of Veterinary Science, Osaka Prefecture University, Sakai, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Tsuyama
authors.forename: E
authors.initials: E
authors.lastname: Yamazaki
authors.forename: T
authors.initials: T
authors.lastname: Tomita
authors.forename: H
authors.initials: H
authors.lastname: Ihara
authors.forename: S
authors.initials: S
authors.lastname: Takenaka
authors.forename: K
authors.initials: K
authors.lastname: Kato
authors.forename: S
authors.initials: S
authors.lastname: Kozaki
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002417:Cattle; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000096:biosynthesis; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000276:immunology* / Q000378:metabolism*; D066298:In Vitro Techniques; D008168:Lung / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D011689:Purkinje Cells / Q000187:drug effects / Q000201:enzymology; D051381:Rats; D012995:Solubility
chemical_list: D000911:Antibodies, Monoclonal; D020030:Nitric Oxide Donors; C110881:S-nitro-N-acetylpenicillamine; D009569:Nitric Oxide; D006162:Guanylate Cyclase; D010396:Penicillamine; D006152:Cyclic GMP