title: Uric acid attenuates nitric oxide production by decreasing the interaction between endothelial nitric oxide synthase and calmodulin in human umbilical vein endothelial cells: a mechanism for uric acid-induced cardiovascular disease development.
abstract: The elevated level of uric acid in the body is associated with increased risk of cardiovascular diseases, which is mediated by endothelial dysfunction. However, its underlying mechanism is not fully understood, although dysregulation of endothelial nitric oxide (NO) production is likely to be involved. Using human umbilical vascular endothelial cells (HUVEC), we explored the molecular mechanism of uric acid on endothelial NO synthase (eNOS) activity and NO production. Although high dose of uric acid (12mg/dl for 24h treatment) significantly decreased eNOS activity and NO production, it did not alter eNOS expression and phosphorylations at eNOS-Ser(1177), eNOS-Thr(495) and eNOS-Ser(114). Under this condition, we also found no alterations in the dimerization and acetylation of eNOS, compared with the control. Furthermore, uric acid did not change the activity of arginase II, an enzyme degrading l-arginine, a substrate of eNOS, and intracellular level of calcium, a cofactor for eNOS activation. We also found that uric acid did not alter xanthine oxidase activity, suggesting no involvement of xanthine oxidase-derived O2(-) production in the observed inhibitory effects. In vitro and in cell coimmunoprecipitation studies, however, revealed that uric acid significantly decreased the interaction between eNOS and calmodulin (CaM), an eNOS activator, although it did not change the intracellular CaM level. Like in HUVEC, uric acid also decreased eNOS-CaM interaction in bovine aortic EC. Finally, uric acid attenuated ionomycin-induced increase in the interaction between eNOS and CaM. This study suggests firstly that uric acid decreased eNOS activity and NO production through reducing the binding between eNOS and CaM in EC. Our result may provide molecular mechanism by which uric acid induces endothelial dysfunction.
authors.affiliation: Department of Molecular Medicine, Ewha Womans University School of Medicine, Seoul 158-710, South Korea.
authors.forename: Jung-Hyun
authors.initials: JH
authors.lastname: Park
authors.forename: Yoon Mi
authors.initials: YM
authors.lastname: Jin
authors.forename: Soojin
authors.initials: S
authors.lastname: Hwang
authors.forename: Du-Hyong
authors.initials: DH
authors.lastname: Cho
authors.forename: Duk-Hee
authors.initials: DH
authors.lastname: Kang
authors.forename: Inho
authors.initials: I
authors.lastname: Jo
mesh_terms: D000818:Animals; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D004347:Drug Interactions; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction; D014527:Uric Acid / Q000494:pharmacology*
chemical_list: D002147:Calmodulin; D014527:Uric Acid; D009569:Nitric Oxide; D015759:Ionomycin; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III