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

J. Biol. Chem., Vol. 279, Issue 10, 9475-9480, March 5, 2004

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 279/10/9475    most recent M307246200v1 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brdlik, C. M. Articles by Crews, C. M. Search for Related Content PubMed PubMed Citation Articles by Brdlik, C. M. Articles by Crews, C. M. Social Bookmarking                      What's this?

A Single Amino Acid Residue Defines the Difference in Ovalicin Sensitivity between Type I and II Methionine Aminopeptidases^*

Cathleen M. Brdlik and Craig M. Crews¶||**

From the Departments of Genetics, Molecular, Cellular, and Developmental Biology, ^¶Pharmacology, and ^||Chemistry, Yale University, New Haven, Connecticut 06520-8103

TNP-470, the first anti-angiogenic small molecule to enter clinical trials, targets methionine aminopeptidase-2 (MetAP-2), a metalloprotease that cleaves the N-terminal methionine of proteins. Previously, biochemical binding, in vivo yeast studies, and structural studies of human methionine aminopeptidase-2 bound to TNP-470 and its analogs fumagillin and ovalicin revealed that these compounds exhibit specificity for MetAP-2 over its family member MetAP-1. To further elucidate the nature of this specificity, we developed a yeast-based screen for human MetAP-2 mutations that confer ovalicin resistance. Of the three resistant alleles, A362T appeared in the majority of clones and was found to be the most resistant to the ovalicin class of inhibitors. Alignment of human MetAP-2 with human MetAP-1, which is naturally ovalicin-resistant, revealed that the analogous residue in MetAP-1 is also a threonine. Mutation of this residue to alanine resulted in an ovalicin-sensitive MetAP-1 allele, demonstrating that an alanine at this position is critical for inhibition by ovalicin. These results provide a molecular explanation for the specificity exhibited by this class of anti-angiogenic agents for MetAP-2 over MetAP-1 and may prove useful in the development of additional MetAP-2-specific therapeutic agents.

Received for publication, July 7, 2003 , and in revised form, November 12, 2003.

^* This work was supported by the National Institutes of Health Grant RO1 CA83049. 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 Supplemental Figs. 1-4.

^** To whom all correspondence should be addressed: Craig M. Crews, 219 Prospect St., New Haven, CT 06520-8103. Tel.: 203-432-9364; Fax: 203-432-6161; E-mail: craig.crews{at}yale.edu.

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

This article has been cited by other articles:

				R. Upadhya, H. S. Zhang, and L. M. Weiss System for Expression of Microsporidian Methionine Amino Peptidase Type 2 (MetAP2) in the Yeast Saccharomyces cerevisiae Antimicrob. Agents Chemother., October 1, 2006; 50(10): 3389 - 3395. [Abstract] [Full Text] [PDF]

				A. Addlagatta and B. W. Matthews Structure of the angiogenesis inhibitor ovalicin bound to its noncognate target, human Type 1 methionine aminopeptidase Protein Sci., August 1, 2006; 15(8): 1842 - 1848. [Abstract] [Full Text] [PDF]

				J.-R. J. Yeh, R. Ju, C. M. Brdlik, W. Zhang, Y. Zhang, M. E. Matyskiela, J. D. Shotwell, and C. M. Crews Targeted gene disruption of methionine aminopeptidase 2 results in an embryonic gastrulation defect and endothelial cell growth arrest PNAS, July 5, 2006; 103(27): 10379 - 10384. [Abstract] [Full Text] [PDF]

				P. J. Didier, J. N. Phillips, D. J. Kuebler, M. Nasr, P. J. Brindley, M. E. Stovall, L. C. Bowers, and E. S. Didier Antimicrosporidial Activities of Fumagillin, TNP-470, Ovalicin, and Ovalicin Derivatives In Vitro and In Vivo. Antimicrob. Agents Chemother., June 1, 2006; 50(6): 2146 - 2155. [Abstract] [Full Text] [PDF]

				S. Ross, C. Giglione, M. Pierre, C. Espagne, and T. Meinnel Functional and Developmental Impact of Cytosolic Protein N-Terminal Methionine Excision in Arabidopsis Plant Physiology, February 1, 2005; 137(2): 623 - 637. [Abstract] [Full Text] [PDF]