JBC Advanced Peptides, Inc.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

A more recent version of this article appeared on July 27, 2001
This Article
Right arrow Full Text (Accepted Manuscript)
Right arrow All Versions of this Article:
276/31/29163    most recent
M101731200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
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 Hubbard, P. A.
Right arrow Articles by Kim, J.-J. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hubbard, P. A.
Right arrow Articles by Kim, J.-J. P.
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?

Papers In Press, published online ahead of print May 22, 2001
J. Biol. Chem, 10.1074/jbc.M101731200
Submitted on February 24, 2001
Revised on May 8, 2001
Accepted on May 22, 2001

NADPH-cytochrome P450 oxidoreductase: structural basis for hydride and electron transfer

Paul A. Hubbard, Anna L. Shen, Rosemary Paschke, Charles B. Kasper, and Jung-Ja P. Kim

Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226

Corresponding Author: jjkim{at}mcw.edu

NADPH-cytochrome P450 oxidoreductase catalyzes transfer of electrons from NADPH, via two flavin cofactors, to various cytochromes P450. The crystal structure of the rat reductase complexed with NADP+ has revealed that nicotinamide access to FAD is blocked by an aromatic residue (W677), which stacks against the re-face of the isoalloxazine ring of the flavin. To investigate the nature of interactions between the nicotinamide, FAD, and W677 during the catalytic cycle, three mutant proteins were studied by crystallography. The first mutant, W677X, has the last two C-terminal residues, W677 and S678, removed; while the second mutant, W677G, retains the C-terminal serine residue. The third mutant has three catalytic residues substituted: S457A, C630A, and D675N. In the W677X and W677G structures, the nicotinamide moiety of NADP+ lies against the FAD isoalloxazine ring with a tilt of approximately 30° between the planes of the two rings. These results, together with the S457A/C630A/D675N structure, allow us to propose a mechanism for hydride transfer regulated by changes in hydrogen bonding and pi-pi interactions between the isoalloxazine ring and either the nicotinamide ring or W677 indole ring. Superimposition of the mutant and wild-type structures shows significant mobility between the two flavin domains of the enzyme. This, together with the high degree of disorder observed in the FMN domain of all three mutant structures, suggests that conformational changes occur during catalysis.


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
 J. Biol. Chem.Home page
R. P. Ilagan, M. Tiso, D. W. Konas, C. Hemann, D. Durra, R. Hille, and D. J. Stuehr
Differences in a Conformational Equilibrium Distinguish Catalysis by the Endothelial and Neuronal Nitric-oxide Synthase Flavoproteins
J. Biol. Chem., July 11, 2008; 283(28): 19603 - 19615.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
A. V. Pandey, P. Kempna, G. Hofer, P. E. Mullis, and C. E. Fluck
Modulation of Human CYP19A1 Activity by Mutant NADPH P450 Oxidoreductase
Mol. Endocrinol., October 1, 2007; 21(10): 2579 - 2595.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. C. Marohnic, S. P. Panda, P. Martasek, and B. S. Masters
Diminished FAD Binding in the Y459H and V492E Antley-Bixler Syndrome Mutants of Human Cytochrome P450 Reductase
J. Biol. Chem., November 24, 2006; 281(47): 35975 - 35982.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. P. Panda, Y. T. Gao, L. J. Roman, P. Martasek, J. C. Salerno, and B. S. S. Masters
The Role of a Conserved Serine Residue within Hydrogen Bonding Distance of FAD in Redox Properties and the Modulation of Catalysis by Ca2+/Calmodulin of Constitutive Nitric-oxide Synthases
J. Biol. Chem., November 10, 2006; 281(45): 34246 - 34257.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. Jachymova, P. Martasek, S. Panda, L. J. Roman, M. Panda, T. M. Shea, Y. Ishimura, J.-J. P. Kim, and B. S. S. Masters
Recruitment of governing elements for electron transfer in the nitric oxide synthase family
PNAS, November 1, 2005; 102(44): 15833 - 15838.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Higashimoto, H. Sakamoto, S. Hayashi, M. Sugishima, K. Fukuyama, G. Palmer, and M. Noguchi
Involvement of NADP(H) in the Interaction between Heme Oxygenase-1 and Cytochrome P450 Reductase
J. Biol. Chem., January 7, 2005; 280(1): 729 - 737.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
E. D. Garcin, C. M. Bruns, S. J. Lloyd, D. J. Hosfield, M. Tiso, R. Gachhui, D. J. Stuehr, J. A. Tainer, and E. D. Getzoff
Structural Basis for Isozyme-specific Regulation of Electron Transfer in Nitric-oxide Synthase
J. Biol. Chem., September 3, 2004; 279(36): 37918 - 37927.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. W. Konas, K. Zhu, M. Sharma, K. S. Aulak, G. W. Brudvig, and D. J. Stuehr
The FAD-shielding Residue Phe1395 Regulates Neuronal Nitric-oxide Synthase Catalysis by Controlling NADP+ Affinity and a Conformational Equilibrium within the Flavoprotein Domain
J. Biol. Chem., August 20, 2004; 279(34): 35412 - 35425.
[Abstract] [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
G. M. Knudsen, C. R. Nishida, S. D. Mooney, and P. R. O. de Montellano
Nitric-oxide Synthase (NOS) Reductase Domain Models Suggest a New Control Element in Endothelial NOS That Attenuates Calmodulin-dependent Activity
J. Biol. Chem., August 22, 2003; 278(34): 31814 - 31824.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. L. Elmore and T. D. Porter
Modification of the Nucleotide Cofactor-binding Site of Cytochrome P-450 Reductase to Enhance Turnover with NADH in Vivo
J. Biol. Chem., December 6, 2002; 277(50): 48960 - 48964.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Adak, M. Sharma, A. L. Meade, and D. J. Stuehr
A conserved flavin-shielding residue regulates NO synthase electron transfer and nicotinamide coenzyme specificity
PNAS, October 15, 2002; 99(21): 13516 - 13521.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Zhang, P. Martasek, R. Paschke, T. Shea, B. S. Siler Masters, and J.-J. P. Kim
Crystal Structure of the FAD/NADPH-binding Domain of Rat Neuronal Nitric-oxide Synthase. COMPARISONS WITH NADPH-CYTOCHROME P450 OXIDOREDUCTASE
J. Biol. Chem., September 28, 2001; 276(40): 37506 - 37513.
[Abstract] [Full Text] [PDF]


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