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Published online on October 16, 2003, 10.1073/pnas.2133986100
PNAS | November 11, 2003 | vol. 100 | no. 23 | 13196-13201


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From The Cover
Biochemistry
An open conformation of mammalian cytochrome P450 2B4 at 1.6-Å resolution

Emily E. Scott * {dagger}, You Ai He *, Michael R. Wester {ddagger}, Mark A. White §, Christopher C. Chin §, James R. Halpert *, Eric F. Johnson {ddagger}, and C. David Stout {dagger} ¶

*Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, TX 77555-1031; Departments of {ddagger}Molecular and Experimental Medicine, MEM-255, and Molecular Biology, MB-8, The Scripps Research Institute, La Jolla, CA 92037; and §Sealy Center for Structural Biology, University of Texas Medical Branch, Galveston, TX 77555-0647

Edited by Minor J. Coon, University of Michigan Medical School, Ann Arbor, MI and approved August 27, 2003 (received for review June 26, 2003)

The xenobiotic metabolizing cytochromes P450 (P450s) are among the most versatile biological catalysts known, but knowledge of the structural basis for their broad substrate specificity has been limited. P450 2B4 has been frequently used as an experimental model for biochemical and biophysical studies of these membrane proteins. A 1.6-Å crystal structure of P450 2B4 reveals a large open cleft that extends from the protein surface directly to the heme iron between the {alpha}-helical and {beta}-sheet domains without perturbing the overall P450 fold. This cleft is primarily formed by helices B' to C and F to G. The conformation of these regions is dramatically different from that of the other structurally defined mammalian P450, 2C5/3LVdH, in which the F to G and B' to C regions encapsulate one side of the active site to produce a closed form of the enzyme. The open conformation of 2B4 is trapped by reversible formation of a homodimer in which the residues between helices F and G of one molecule partially fill the open cleft of a symmetry-related molecule, and an intermolecular coordinate bond occurs between H226 and the heme iron. This dimer is observed both in solution and in the crystal. Differences between the structures of 2C5 and 2B4 suggest that defined regions of xenobiotic metabolizing P450s may adopt a substantial range of energetically accessible conformations without perturbing the overall fold. This conformational flexibility is likely to facilitate substrate access, metabolic versatility, and product egress.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: P450, cytochrome P450; SRS, substrate recognition site.

Data deposition: The atomic coordinates and structure factors have been deposited in the Protein Data Bank, www.rcsb.org (PDB ID code 1PO5).

See Commentary on page 13121.

{dagger} To whom correspondence should be addressed at: Department of Pharmacology and Toxicology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-1031. E-mail: eescott{at}x-ray.utmb.edu or dave{at}scripps.edu.


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Related Commentary in PNAS:

Cytochrome P450 flexibility
Thomas L. Poulos
PNAS 2003 100: 13121-13122. [Extract] [Full Text]  



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