HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 10, 9088-9096, March 11, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/10/9088    most recent M413042200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Matsuura, K. Articles by Waterman, M. R. Search for Related Content PubMed PubMed Citation Articles by Matsuura, K. Articles by Waterman, M. R. Social Bookmarking                      What's this?

Structural Diversities of Active Site in Clinical Azole-bound Forms between Sterol 14-Demethylases (CYP51s) from Human and Mycobacterium tuberculosis^*

Koji Matsuura, Shiro Yoshioka¶||, Takehiko Tosha**, Hiroshi Hori, Koichiro Ishimori, Teizo Kitagawa**, Isao Morishima, Norio Kagawa¶, and Michael R. Waterman¶

From the Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan, ^¶Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146, ^**Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Okazaki 444-8787, Japan, and Graduate School of Engineering Science, Osaka University, Toyonaka 560-8531, Japan

To gain insights into the molecular basis of the design for the selective azole anti-fungals, we compared the binding properties of azole-based inhibitors for cytochrome P450 sterol 14-demethylase (CYP51) from human (HuCYP51) and Mycobacterium tuberculosis (MtCYP51). Spectroscopic titration of azoles to the CYP51s revealed that HuCYP51 has higher affinity for ketoconazole (KET), an azole derivative that has long lipophilic groups, than MtCYP51, but the affinity for fluconazole (FLU), which is a member of the anti-fungal armamentarium, was lower in HuCYP51. The affinity for 4-phenylimidazole (4-PhIm) to MtCYP51 was quite low compared with that to HuCYP51. In the resonance Raman spectra for HuCYP51, the FLU binding induced only minor spectral changes, whereas the prominent high frequency shift of the bending mode of the heme vinyl group was detected in the KET- or 4-PhIm-bound forms. On the other hand, the bending mode of the heme propionate group for the FLU-bound form of MtCYP51 was shifted to high frequency as found for the KET-bound form, but that for 4-PhIm was shifted to low frequency. The EPR spectra for 4-PhIm-bound MtCYP51 and FLU-bound HuCYP51 gave multiple g values, showing heterogeneous binding of the azoles, whereas the single g_x and g_z values were observed for other azole-bound forms. Together with the alignment of the amino acid sequence, these spectroscopic differences suggest that the region between the B' and C helices, particularly the hydrophobicity of the C helix, in CYP51s plays primary roles in determining strength of interactions with azoles; this differentiates the binding specificity of azoles to CYP51s.

Received for publication, November 18, 2004 , and in revised form, December 14, 2004.

^* This work was supported by Grants-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology in Japan (12002008 to I. M., 13680741 to H. H., and 14658217 to K. I.) and Grants GM37942 and GM67871 from the National Institutes of Health (to M. R. W.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Present address: Research Center for Advanced Carbon Materials, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8565, Japan.

^|| Present address: Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Okazaki, 444-8787, Japan.

To whom correspondence should be addressed. Tel.: 81-75-383-2536; Fax: 81-75-383-2541; E-mail: koichiro{at}scl.kyoto-u.ac.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				K. J. McLean, P. Carroll, D. G. Lewis, A. J. Dunford, H. E. Seward, R. Neeli, M. R. Cheesman, L. Marsollier, P. Douglas, W. E. Smith, et al. Characterization of Active Site Structure in CYP121: A CYTOCHROME P450 ESSENTIAL FOR VIABILITY OF MYCOBACTERIUM TUBERCULOSIS H37Rv J. Biol. Chem., November 28, 2008; 283(48): 33406 - 33416. [Abstract] [Full Text] [PDF]

				S. Ekins, C. Chang, S. Mani, M. D. Krasowski, E. J. Reschly, M. Iyer, V. Kholodovych, N. Ai, W. J. Welsh, M. Sinz, et al. Human Pregnane X Receptor Antagonists and Agonists Define Molecular Requirements for Different Binding Sites Mol. Pharmacol., September 1, 2007; 72(3): 592 - 603. [Abstract] [Full Text] [PDF]

				A. J. Dunford, K. J. McLean, M. Sabri, H. E. Seward, D. J. Heyes, N. S. Scrutton, and A. W. Munro Rapid P450 Heme Iron Reduction by Laser Photoexcitation of Mycobacterium tuberculosis CYP121 and CYP51B1: ANALYSIS OF CO COMPLEXATION REACTIONS AND REVERSIBILITY OF THE P450/P420 EQUILIBRIUM J. Biol. Chem., August 24, 2007; 282(34): 24816 - 24824. [Abstract] [Full Text] [PDF]

				S. Ekins, D. C. Mankowski, D. J. Hoover, M. P. Lawton, J. L. Treadway, and H. J. Harwood Jr. Three-Dimensional Quantitative Structure-Activity Relationship Analysis of Human CYP51 Inhibitors Drug Metab. Dispos., March 1, 2007; 35(3): 493 - 500. [Abstract] [Full Text] [PDF]

				A. Verras, A. Alian, and P. R.O. d. Montellano Cytochrome P450 active site plasticity: attenuation of imidazole binding in cytochrome P450cam by an L244A mutation Protein Eng. Des. Sel., November 1, 2006; 19(11): 491 - 496. [Abstract] [Full Text] [PDF]

				M. P. Pietila, P. K. Vohra, B. Sanyal, N. L. Wengenack, S. Raghavakaimal, and C. F. Thomas Jr. Cloning and Characterization of CYP51 from Mycobacterium avium Am. J. Respir. Cell Mol. Biol., August 1, 2006; 35(2): 236 - 242. [Abstract] [Full Text] [PDF]

				Y. Zhao, M. A. White, B. K. Muralidhara, L. Sun, J. R. Halpert, and C. D. Stout Structure of Microsomal Cytochrome P450 2B4 Complexed with the Antifungal Drug Bifonazole: INSIGHT INTO P450 CONFORMATIONAL PLASTICITY AND MEMBRANE INTERACTION J. Biol. Chem., March 3, 2006; 281(9): 5973 - 5981. [Abstract] [Full Text] [PDF]