title: Role of PKC-ζ in NADPH oxidase-PKCα-Giα axis dependent inhibition of β-adrenergic response by U46619 in pulmonary artery smooth muscle cells.
abstract: Treatment of bovine pulmonary artery smooth muscle cells (BPASMCs) with U46619 attenuated isoproterenol caused stimulation of adenyl cyclase activity and cAMP production. Pretreatment with SQ29548 (Tp receptor antagonist), apocynin (NADPH oxidase inhibitor) and Go6976 (PKC-α inhibitor) eliminated U46619 caused attenuation of isoproterenol stimulated adenyl cyclase activity. Pretreatment with SQ29548 and apocynin prevented U46619 induced increase in NADPH oxidase activity, PKC-α activity and Giα phosphorylation. However, pretreatment with CZI, a PKC-ζ inhibitor, markedly, but not completely, inhibited U46619 induced increase in NADPH oxidase activity, PKC-α activity, Giα phosphorylation and also significantly eliminated U46619 caused attenuation of isoproterenol stimulated adenyl cyclase activity. Pretreatment with Go6976 inhibited U46619 induced increase in Giα phosphorylation, but not PKC-ζ activity and NADPH oxidase activity. Pretreatment with pertussis toxin eliminated U46619 caused attenuation of isoproterenol stimulated adenyl cyclase activity without any discernible change in PKC-ζ, NADPH oxidase and PKC-α activities. Transfection of the cells with Tp, PKC-ζ and PKC-α siRNA duplexes corroborate the findings observed with their respective pharmacological inhibitors on the responses produced by U46619. Taken together, we suggest involvement of PKC-ζ in U46619 caused attenuation of isoproterenol stimulated β-adrenergic response, which is regulated by NADPH oxidase-PKCα-Giα axis in pulmonary artery smooth muscle cells.
authors.affiliation: Department of Biochemistry and Biophysics, University of Kalyani, Nadia 741235, West Bengal, India. Electronic address: sajal_chakraborti@yahoo.com.
authors.forename: Sajal
authors.initials: S
authors.lastname: Chakraborti
authors.forename: Soumitra
authors.initials: S
authors.lastname: Roy
authors.forename: Amritlal
authors.initials: A
authors.lastname: Mandal
authors.forename: Animesh
authors.initials: A
authors.lastname: Chowdhury
authors.forename: Tapati
authors.initials: T
authors.lastname: Chakraborti
mesh_terms: D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology*; D000262:Adenylyl Cyclases / Q000378:metabolism; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha / Q000378:metabolism*; D011651:Pulmonary Artery / Q000166:cytology; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D000099221:Protein Kinase C zeta
chemical_list: D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D000262:Adenylyl Cyclases; D000319:Adrenergic beta-Antagonists; D000242:Cyclic AMP; D004791:Enzyme Inhibitors; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D007527:Isoenzymes; D007545:Isoproterenol; D019255:NADPH Oxidases; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D011943:Receptors, Adrenergic, beta; D000099221:Protein Kinase C zeta
keywords: Adenyl cyclase; G(i)α; PKC-α; PKC-ζ; U46619; cAMP