Ca2+/calmodulin-dependent nitric oxide synthase activity in the human cervix carcinoma cell line ME-180
- 15 January 1993
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 289 (2) , 357-361
- https://doi.org/10.1042/bj2890357
Abstract
We show here that the human cervix carcinoma cell line ME-180 expresses a constitutive nitric oxide (NO) synthase, as demonstrated by formation of [3H]citrulline and nitrite. The enzyme is dependent on tetrahydrobiopterin, NADPH, flavins and Ca2+/calmodulin. Enzyme activity is located in the cytosol rather than in the membrane fraction and can be inhibited by NG-monomethyl-L-arginine (NMMA). An antiserum to NO synthase purified from porcine cerebellum inhibited the enzyme activity. ME-180 cells released NO, as was shown by stimulation of guanylate cyclase (EC 4.6.1.2) in RFL-6 detector cells; this release was stimulated 8-fold by the Ca2+ ionophore A23187 and 2-fold by increasing the intracellular tetrahydrobiopterin levels with cytokines. This is the first characterization of a Ca2+/calmodulin-dependent NO synthase activity in human epithelial-type tumour cells.Keywords
This publication has 35 references indexed in Scilit:
- Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells.Proceedings of the National Academy of Sciences, 1991
- Human colorectal adenocarcinoma cells: differential nitric oxide synthesis determines their ability to aggregate platelets.1991
- Purification and characterization of the cytokine-induced macrophage nitric oxide synthase: an FAD- and FMN-containing flavoprotein.Proceedings of the National Academy of Sciences, 1991
- Nitric oxide: physiology, pathophysiology, and pharmacology.1991
- Nitric oxide: an endogenous modulator of leukocyte adhesion.Proceedings of the National Academy of Sciences, 1991
- Calmodulin-dependent endothelium-derived relaxing factor/nitric oxide synthase activity is present in the particulate and cytosolic fractions of bovine aortic endothelial cells.Proceedings of the National Academy of Sciences, 1991
- Calmodulin-independent nitric oxide synthase from rat polymorphonuclear neutrophilsJournal of Biological Chemistry, 1991
- Purification of a Ca2+/calmodulin‐dependent nitric oxide synthase from porcine cerebellumFEBS Letters, 1990
- 7-Substituted pterins: Formation during phenylalanine hydroxylation in the absence of dehydrataseBiochemical and Biophysical Research Communications, 1990
- An L-arginine/nitric oxide pathway present in human platelets regulates aggregation.Proceedings of the National Academy of Sciences, 1990