PMID: 9173876Apr 1, 1997Paper

Characterization of bovine endothelial nitric oxide synthase as a homodimer with down-regulated uncoupled NADPH oxidase activity: tetrahydrobiopterin binding kinetics and role of haem in dimerization

The Biochemical Journal
B M ListB Mayer

Abstract

The fatty-acylation-deficient bovine endothelial NO synthase (eNOS) mutant (Gly-2 to Ala-2, G2AeNOS) was purified from a baculovirus overexpression system. The purified protein was soluble and highly active (0.2-0.7 micromol of l-citrulline. mg-1.min-1), contained 0. 77+/-0.01 equivalent of haem per subunit, showed a Soret maximum at 396 nm, and exhibited only minor uncoupling of NADPH oxidation in the absence of l-arginine or tetrahydrobiopterin. Radioligand binding studies revealed KD values of 147+/-24.1 nM and 52+/-9.2 nM for specific binding of tetrahydrobiopterin in the absence and presence of 0.1 mM l-arginine respectively. The positive co-operative effect of l-arginine was due to a pronounced decrease in the rate of tetrahydrobiopterin dissociation (from 1.6+/-0.5 to 0. 3+/-0.1 min-1). Low-temperature SDS gel electrophoresis showed that approx. 80% of the protein migrated as haem-containing dimer after preincubation with l-arginine and tetrahydrobiopterin. Gel-filtration chromatography yielded one peak with a Stokes radius of 6.8+/-0.04 nm, corresponding to a hydrodynamic volume of 1. 32x10(-24) m3, whereas haem-deficient preparations (approx. 0.3 equivalent per subunit) contained an additional protein species with a hy...Continue Reading

Citations

Feb 27, 2013·IUBMB Life·Jesús Tejero, Dennis Stuehr
Sep 16, 2015·Current Hypertension Reports·Eugenia GkaliagkousiStella Douma
Oct 4, 2006·Biochimica Et Biophysica Acta·Antonius C F Gorren, Bernd Mayer
Jul 11, 2013·Oral Diseases·M P Hezel, E Weitzberg
Apr 22, 2003·Progress in Cardiovascular Diseases·Xing Li WangJian Wang
May 28, 2013·Free Radical Biology & Medicine·Jennifer WhitsettJeannette Vasquez-Vivar
Mar 14, 2002·Nitric Oxide : Biology and Chemistry·Andrew J Maxwell
Aug 24, 2000·The Journal of Biological Chemistry·B HemmensB Mayer
Jun 6, 2002·The Journal of Biological Chemistry·Koustubh PandaDennis J Stuehr
Sep 1, 2004·Histochemistry and Cell Biology·R M Touyz, E L Schiffrin
Dec 11, 2002·The Journal of Pathology·Ester W J A AlbrechtHarry van Goor
Apr 7, 2007·Arteriosclerosis, Thrombosis, and Vascular Biology·Lila MayahiAroon D Hingorani
Oct 13, 2010·Circulation. Cardiovascular Genetics·Lila MayahiAroon D Hingorani
Sep 20, 2007·Journal of Cardiovascular Pharmacology·An L Moens, David A Kass
Mar 20, 2004·Expert Review of Cardiovascular Therapy·Rhian M Touyz
Aug 5, 1998·Proceedings of the National Academy of Sciences of the United States of America·J Vásquez-VivarK A Pritchard
Oct 20, 2005·Dalton Transactions : an International Journal of Inorganic Chemistry·Dennis J StuehrRuss Hille
Sep 6, 2011·European Heart Journal·Ulrich Förstermann, William C Sessa
Mar 29, 2014·Pharmacological Reviews·Marie BillaudBrant E Isakson
May 27, 2005·American Journal of Physiology. Cell Physiology·Davin JagnandanDavid Fulton
Mar 13, 2001·Journal of Cardiovascular Pharmacology·T A BakerZ S Katusic
Mar 30, 2001·The Journal of Biological Chemistry·D StuehrG M Rosen
Apr 5, 2000·The Journal of Biological Chemistry·W WangR L Danner
Aug 22, 2000·The Journal of Biological Chemistry·A T BenderY Osawa
Mar 13, 1999·The Journal of Biological Chemistry·R HellerU Till
Oct 3, 1999·The Journal of Biological Chemistry·M TeublK Schmidt
Mar 4, 2000·Proceedings of the National Academy of Sciences of the United States of America·K McMillanJ J Devlin

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