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

J. Biol. Chem., Vol. 282, Issue 27, 19948-19957, July 6, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/27/19948    most recent M701480200v1 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 Google Scholar Google Scholar Articles by Miallau, L. Articles by Leonard, G. A. Search for Related Content PubMed PubMed Citation Articles by Miallau, L. Articles by Leonard, G. A. Social Bookmarking                      What's this?

Structures of Staphylococcus aureus D-Tagatose-6-phosphate Kinase Implicate Domain Motions in Specificity and Mechanism^*

Linda Miallau, William N. Hunter¹, Sean M. McSweeney, and Gordon A. Leonard²

From the Macromolecular Crystallography Group, European Synchrotron Radiation Facility, 38043 Grenoble, France and Division of Biological Chemistry and Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom

High resolution structures of Staphylococcus aureus D-tagatose-6-phosphate kinase (LacC) in two crystal forms are herein reported. The structures define LacC in apoform, in binary complexes with ADP or the co-factor analogue AMP-PNP, and in a ternary complex with AMP-PNP and D-tagatose-6-phosphate. The tertiary structure of the LacC monomer, which is closely related to other members of the pfkB subfamily of carbohydrate kinases, is composed of a large / core domain and a smaller, largely "lid." Four extended polypeptide segments connect these two domains. Dimerization of LacC occurs via interactions between lid domains, which come together to form a -clasp structure. Residues from both subunits contribute to substrate binding. LacC adopts a closed structure required for phosphoryl transfer only when both substrate and co-factor are bound. A reaction mechanism similar to that used by other phosphoryl transferases is proposed, although unusually, when both substrate and co-factor are bound to the enzyme two Mg²⁺ ions are observed in the active site. A new motif of amino acid sequence conservation common to the pfkB subfamily of carbohydrate kinases is identified.

Received for publication, February 20, 2007 , and in revised form, April 6, 2007.

The atomic coordinates and structure factors (code 2JGV and 2JG1) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* 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.

¹ Supported by the Wellcome Trust and the Biotechnology and Biological Sciences Research Council (Structural Proteomics of Rational Targets).

² To whom correspondence should be addressed: Macomolecular Crystallography Group, European Synchrotron Radiation Facility, 6 Rue Jules Horowitz, 38043 Grenoble Cedex, France. Tel.: 33-4-76-88-23-94; Fax: 33-4-76-88-29-04; E-mail: leonard{at}esrf.fr.

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