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Mechanism of Inactivation of Neuronal Nitric Oxide Synthase by Nω-Allyl-L-arginine
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Zhang, H. Q.; Dixon, R. P.; Marletta, M. A.; Nikolic, D.; Van Breemen, R.; Silverman, R. B. Mechanism of Inactivation of Neuronal Nitric Oxide Synthase by Nω-Allyl-L-arginine. J. Am Chem. Soc. 1997, 119, in press.
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The determination of enzyme inhibitor constants
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The competitive reversible inhibition of the isoforms by Nω-propyl-L-arginine was studied under initial rate conditions with the hemoglobin assay as described in ref 10. The concentrations of L-arginine used were 3, 4, 6, 10, and 15 μM for nNOS; 10, 20, 30, and 60 μM for iNOS; and 2, 4, 8, and 15 μM for eNOS, The concentrations of Nμ-propyl-L-arginine used were 0.05, 0.1, 0.15, and 0.2 μM for nNOS; 50, 100, 150, and 200 μM for iNOS; and 10, 20, 30, and 40 μM for eNOS. Data were analyzed by the methods of Dixon (Dixon, M. The determination of enzyme inhibitor constants. Biochem. J. 1953, 55, 170-171) and Cornish-Bowden (Cornish-Bowden, A. Fundamentals of Enzyme Kinetics; Portland Press: London, 1995; pp 102-108).
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The competitive reversible inhibition of the isoforms by Nω-propyl-L-arginine was studied under initial rate conditions with the hemoglobin assay as described in ref 10. The concentrations of L-arginine used were 3, 4, 6, 10, and 15 μM for nNOS; 10, 20, 30, and 60 μM for iNOS; and 2, 4, 8, and 15 μM for eNOS, The concentrations of Nμ-propyl-L-arginine used were 0.05, 0.1, 0.15, and 0.2 μM for nNOS; 50, 100, 150, and 200 μM for iNOS; and 10, 20, 30, and 40 μM for eNOS. Data were analyzed by the methods of Dixon (Dixon, M. The determination of enzyme inhibitor constants. Biochem. J. 1953, 55, 170-171) and Cornish-Bowden (Cornish-Bowden, A. Fundamentals of Enzyme Kinetics; Portland Press: London, 1995; pp 102-108).
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Characterization of Bovine Endothelial Nitric Oxide Synthase Expressed in E. coli
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Martasek, P.; Liu, Q.; Liu, J.; Roman, L. J.; Gross, S. S.; Sessa, W. C.; Masters, B. S. S. Characterization of Bovine Endothelial Nitric Oxide Synthase Expressed in E. coli. Biochem. Biophys. Res. Commun. 1996, 219, 359-365.
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