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

J. Biol. Chem., Vol. 283, Issue 22, 15152-15159, May 30, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/22/15152    most recent M801145200v1 Alert me when this article is cited 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 Google Scholar Articles by Eddine, A. N. Articles by Podust, L. M. PubMed PubMed Citation Articles by Eddine, A. N. Articles by Podust, L. M. Related Collections Papers Of The Week Social Bookmarking                      What's this?

X-ray Structure of 4,4'-Dihydroxybenzophenone Mimicking Sterol Substrate in the Active Site of Sterol 14-Demethylase (CYP51)^*

Ali Nasser Eddine, Jens P. von Kries, Mikhail V. Podust^¶, Thulasi Warrier, Stefan H. E. Kaufmann, and Larissa M. Podust^¶1

From the Max-Planck-Institute for Infection Biology, Berlin, 10117, Germany, the Screening Unit, Leibniz-Institute for Molecular Pharmacology (FMP), Berlin, 13125, Germany, and the ^¶Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158

A universal step in the biosynthesis of membrane sterols and steroid hormones is the oxidative removal of the 14-methyl group from sterol precursors by sterol 14-demethylase (CYP51). This enzyme is a primary target in treatment of fungal infections in organisms ranging from humans to plants, and development of more potent and selective CYP51 inhibitors is an important biological objective. Our continuing interest in structural aspects of substrate and inhibitor recognition in CYP51 led us to determine (to a resolution of 1.95Å) the structure of CYP51 from Mycobacterium tuberculosis (CYP51_Mt) co-crystallized with 4,4'-dihydroxybenzophenone (DHBP), a small organic molecule previously identified among top type I binding hits in a library screened against CYP51_Mt. The newly determined CYP51_Mt-DHBP structure is the most complete to date and is an improved template for three-dimensional modeling of CYP51 enzymes from fungal and prokaryotic pathogens. The structure demonstrates the induction of conformational fit of the flexible protein regions and the interactions of conserved Phe-89 essential for both fungal drug resistance and catalytic function, which were obscure in the previously characterized CYP51_Mt-estriol complex. DHBP represents a benzophenone scaffold binding in the CYP51 active site via a type I mechanism, suggesting (i) a possible new class of CYP51 inhibitors targeting flexible regions, (ii) an alternative catalytic function for bacterial CYP51 enzymes, and (iii) a potential for hydroxybenzophenones, widely distributed in the environment, to interfere with sterol biosynthesis. Finally, we show the inhibition of M. tuberculosis growth by DHBP in a mouse macrophage model.

Received for publication, February 12, 2008 , and in revised form, March 13, 2008.

The atomic coordinates and structure factors (code 2VKU) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* This work was supported, in whole or in part, by National Institutes of Health RO1 Grant GM078553 (to L. M. P.). This work was also supported by U.S. Civilian Research and Development Foundation Grant RUB1-2806-MO-06 (to L. M. P.) and by the X-Mtb consortium Bundesministerium fuer Bildung und Forschung/Projekttraeger Juelich Grants BIO/0312992A (to J. P. K.) and 0312992C (to S. H. E. K. and A. N. E.). 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.

The on-line version of this article (available at http://www.jbc.org) contains a supplemental figure.

This article was selected as a Paper of the Week.

¹ To whom correspondence should be addressed: Dept. of Pharmaceutical Chemistry, University of California, 600 16th St., San Francisco, CA 54158-2280. Fax: 415-502-4728; E-mail: larissa.podust{at}ucsf.edu.

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