JBC Avanti Polar Lipids

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M006858200 on October 18, 2000

J. Biol. Chem., Vol. 276, Issue 2, 1233-1243, January 12, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/2/1233    most recent
M006858200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Santolini, J.
Right arrow Articles by Stuehr, D. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Santolini, J.
Right arrow Articles by Stuehr, D. J.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

A Kinetic Simulation Model That Describes Catalysis and Regulation in Nitric-oxide Synthase*

Jérôme Santolini, Subrata AdakDagger , Christine M. L. Curran, and Dennis J. Stuehr§

From the Department of Immunology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195

After initiating NO synthesis a majority of neuronal NO synthase (nNOS) quickly partitions into a ferrous heme-NO complex. This down-regulates activity and increases enzyme Km,O2. To understand this process, we developed a 10-step kinetic model in which the ferric heme-NO enzyme forms as the immediate product of catalysis, and then partitions between NO dissociation versus reduction to a ferrous heme-NO complex. Rate constants used for the model were derived from recent literature or were determined here. Computer simulations of the model precisely described both pre-steady and steady-state features of nNOS catalysis, including NADPH consumption and NO production, buildup of a heme-NO complex, changes between pre-steady and steady-state rates, and the change in enzyme Km,O2 in the presence or absence of NO synthesis. The model also correctly simulated the catalytic features of nNOS mutants W409F and W409Y, which are hyperactive and display less heme-NO complex formation in the steady state. Model simulations showed how the rate of heme reduction influences several features of nNOS catalysis, including populations of NO-bound versus NO-free enzyme in the steady state and the rate of NO synthesis. The simulation predicts that there is an optimum rate of heme reduction that is close to the measured rate in nNOS. Ratio between NADPH consumption and NO synthesis is also predicted to increase with faster heme reduction. Our kinetic model is an accurate and versatile tool for understanding catalytic behavior and will provide new perspectives on NOS regulation.

* This work was supported by National Institutes of Health Grant GM51491.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Fellow of the American Heart Association.

§ To whom correspondence should be addressed: Immunology NB-3, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Ave., Cleveland, OH 44195. Tel.: 216-445-6950; Fax: 216-444-9329; E-mail: stuehrd@ccf.org.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
M. M. Haque, K. Panda, J. Tejero, K. S. Aulak, M. A. Fadlalla, A. T. Mustovich, and D. J. Stuehr
A connecting hinge represses the activity of endothelial nitric oxide synthase
PNAS, May 29, 2007; 104(22): 9254 - 9259.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Marechal, T. A. Mattioli, D. J. Stuehr, and J. Santolini
Activation of Peroxynitrite by Inducible Nitric-oxide Synthase: A DIRECT SOURCE OF NITRATIVE STRESS
J. Biol. Chem., May 11, 2007; 282(19): 14101 - 14112.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Tiso, D. W. Konas, K. Panda, E. D. Garcin, M. Sharma, E. D. Getzoff, and D. J. Stuehr
C-terminal Tail Residue Arg1400 Enables NADPH to Regulate Electron Transfer in Neuronal Nitric-oxide Synthase
J. Biol. Chem., November 25, 2005; 280(47): 39208 - 39219.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. J. Stuehr, J. Santolini, Z.-Q. Wang, C.-C. Wei, and S. Adak
Update on Mechanism and Catalytic Regulation in the NO Synthases
J. Biol. Chem., August 27, 2004; 279(35): 36167 - 36170.
[Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Panda, S. Adak, D. Konas, M. Sharma, and D. J. Stuehr
A Conserved Aspartate (Asp-1393) Regulates NADPH Reduction of Neuronal Nitric-oxide Synthase: IMPLICATIONS FOR CATALYSIS
J. Biol. Chem., April 30, 2004; 279(18): 18323 - 18333.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Z.-Q. Wang, C.-C. Wei, M. Sharma, K. Pant, B. R. Crane, and D. J. Stuehr
A Conserved Val to Ile Switch near the Heme Pocket of Animal and Bacterial Nitric-oxide Synthases Helps Determine Their Distinct Catalytic Profiles
J. Biol. Chem., April 30, 2004; 279(18): 19018 - 19025.
[Abstract] [Full Text] [PDF]

Home page
 Sci SignalHome page
S. J. Vayttaden and U. S. Bhalla
Developing Complex Signaling Models Using GENESIS/Kinetikit
Sci. Signal., February 10, 2004; 2004(219): pl4 - pl4.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Panda, S. Adak, K. S. Aulak, J. Santolini, J. F. McDonald, and D. J. Stuehr
Distinct Influence of N-terminal Elements on Neuronal Nitric-oxide Synthase Structure and Catalysis
J. Biol. Chem., September 26, 2003; 278(39): 37122 - 37131.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. H. Craig, S. K. Chapman, and S. Daff
Calmodulin Activates Electron Transfer through Neuronal Nitric-oxide Synthase Reductase Domain by Releasing an NADPH-dependent Conformational Lock
J. Biol. Chem., September 6, 2002; 277(37): 33987 - 33994.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Adak, K. S. Aulak, and D. J. Stuehr
Direct Evidence for Nitric Oxide Production by a Nitric-oxide Synthase-like Protein from Bacillus subtilis
J. Biol. Chem., May 3, 2002; 277(18): 16167 - 16171.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Z.-Q. Wang, C.-C. Wei, and D. J. Stuehr
A Conserved Tryptophan 457 Modulates the Kinetics and Extent of N-Hydroxy-L-Arginine Oxidation by Inducible Nitric-oxide Synthase
J. Biol. Chem., April 5, 2002; 277(15): 12830 - 12837.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Kobayashi, S. Tagawa, S. Daff, I. Sagami, and T. Shimizu
Rapid Calmodulin-dependent Interdomain Electron Transfer in Neuronal Nitric-oxide Synthase Measured by Pulse Radiolysis
J. Biol. Chem., October 19, 2001; 276(43): 39864 - 39871.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Adak, K. S. Aulak, and D. J. Stuehr
Chimeras of Nitric-oxide Synthase Types I and III Establish Fundamental Correlates between Heme Reduction, Heme-NO Complex Formation, and Catalytic Activity
J. Biol. Chem., June 22, 2001; 276(26): 23246 - 23252.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Couture, S. Adak, D. J. Stuehr, and D. L. Rousseau
Regulation of the Properties of the Heme-NO Complexes in Nitric-oxide Synthase by Hydrogen Bonding to the Proximal Cysteine
J. Biol. Chem., October 5, 2001; 276(41): 38280 - 38288.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Santolini, A. L. Meade, and D. J. Stuehr
Differences in Three Kinetic Parameters Underpin the Unique Catalytic Profiles of Nitric-oxide Synthases I, II, and III
J. Biol. Chem., December 21, 2001; 276(52): 48887 - 48898.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. Stuehr, S. Pou, and G. M. Rosen
Oxygen Reduction by Nitric-oxide Synthases
J. Biol. Chem., April 27, 2001; 276(18): 14533 - 14536.
[Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.