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

J. Biol. Chem., Vol. 277, Issue 26, 23596-23603, June 28, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/26/23596    most recent M202288200v1 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 Boukhvalova, M. Articles by Stewart, R. C. Search for Related Content PubMed PubMed Citation Articles by Boukhvalova, M. Articles by Stewart, R. C. Social Bookmarking                      What's this?

CheA Kinase and Chemoreceptor Interaction Surfaces on CheW^*

Marina Boukhvalova, Ricaele VanBruggen, and Richard C. Stewart

From the Department of Cell Biology and Molecular Genetics and Graduate Program in Molecular and Cellular Biology, University of Maryland, College Park, Maryland 20742

Chemotactic responses of Escherichia coli to aspartic acid are initiated by a ternary protein complex composed of Tar (chemoreceptor), CheA (kinase), and CheW (a coupling protein that binds to both Tar and CheA and links their activities). We used a genetic selection based on the yeast two-hybrid assay to identify nine cheW point mutations that specifically disrupted CheW interaction with CheA but not with Tar. We sequenced these single point mutants and purified four of the mutant CheW proteins for detailed biochemical characterizations that demonstrated the weakened affinity of the mutant CheW proteins for CheA, but not for Tar. In the three-dimensional structure of CheW, the positions affected by these mutations cluster on one face of the protein, defining a potential binding interface for interaction of CheW with CheA. We used a similar two-hybrid approach to identify four mutation sites that disrupted CheW binding to Tar. Mapping of these "Tar-sensitive" mutation sites and those from previous suppressor analysis onto the structure of CheW defined an extended surface on a face of the protein that is adjacent to the CheA-binding surface and that may serve as an interface for CheW binding to Tar.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				P. Mowery, J. B. Ostler, and J. S. Parkinson Different Signaling Roles of Two Conserved Residues in the Cytoplasmic Hairpin Tip of Tsr, the Escherichia coli Serine Chemoreceptor J. Bacteriol., December 15, 2008; 190(24): 8065 - 8074. [Abstract] [Full Text] [PDF]

				J. Zhao and J. S. Parkinson Cysteine-Scanning Analysis of the Chemoreceptor-Coupling Domain of the Escherichia coli Chemotaxis Signaling Kinase CheA. J. Bacteriol., June 1, 2006; 188(12): 4321 - 4330. [Abstract] [Full Text] [PDF]

				S. R. Thompson, G. H. Wadhams, and J. P. Armitage The positioning of cytoplasmic protein clusters in bacteria PNAS, May 23, 2006; 103(21): 8209 - 8214. [Abstract] [Full Text] [PDF]

				J. Zhao and J. S. Parkinson Mutational Analysis of the Chemoreceptor-Coupling Domain of the Escherichia coli Chemotaxis Signaling Kinase CheA J. Bacteriol., May 1, 2006; 188(9): 3299 - 3307. [Abstract] [Full Text] [PDF]

				N. R. Francis, P. M. Wolanin, J. B. Stock, D. J. DeRosier, and D. R. Thomas From the Cover: Three-dimensional structure and organization of a receptor/signaling complex PNAS, December 14, 2004; 101(50): 17480 - 17485. [Abstract] [Full Text] [PDF]

				G. Alexandre and I. B. Zhulin Different Evolutionary Constraints on Chemotaxis Proteins CheW and CheY Revealed by Heterologous Expression Studies and Protein Sequence Analysis J. Bacteriol., January 15, 2003; 185(2): 544 - 552. [Abstract] [Full Text] [PDF]