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

J. Biol. Chem., Vol. 280, Issue 9, 7769-7776, March 4, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/9/7769    most recent M411736200v1 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 Lieser, S. A. Articles by Adams, J. A. Search for Related Content PubMed PubMed Citation Articles by Lieser, S. A. Articles by Adams, J. A. Social Bookmarking                      What's this?

Phosphoryl Transfer Step in the C-terminal Src Kinase Controls Src Recognition^*

Scot A. Lieser**, Caitlin Shindler, Brandon E. Aubol¶, Sungsoo Lee||, Gongqin Sun||, and Joseph A. Adams

From the Department of Pharmacology, University of California at San Diego, La Jolla, California 92093-0728, Department of Chemistry & Biochemistry, University of California at San Diego, La Jolla, California 92093-0728, and ^||Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881

All members of the Src family of nonreceptor protein tyrosine kinases are phosphorylated and subsequently down-regulated by the C-terminal Src kinase, Csk. Although the recognition of Src protein substrates is essential for a diverse set of signaling events linked to cellular growth and differentiation, the factors controlling this critical protein-protein interaction are not well known. To understand how Csk recognizes Src, the chemical/physical events that modulate apparent substrate affinity and turnover were investigated. Src is phosphorylated in a biphasic manner in rapid quench flow experiments, suggesting that the phosphoryl transfer step is fast and highly favorable and does not limit overall turnover. As opposed to other kinase-substrate pairs, turnover is not limited by the physical release of ADP based on stopped-flow fluorescence and catalytic trapping experiments, suggesting that other steps control net phosphorylation. The K_d for Src is considerably larger than the K_m based on single turnover kinetic and equilibrium sedimentation experiments. Taken together, the data are consistent with a mechanism whereby Csk achieves a low K_m for the substrate Src, not by stabilizing protein-protein interactions but rather by facilitating a fast phosphoryl transfer step. In this manner, the phosphoryl transfer step functions as a chemical clamp facilitating substrate recognition.

Received for publication, October 15, 2004 , and in revised form, December 9, 2004.

^* This work was supported by National Institutes of Health Grants GM 68168 (to J. A. A.) and 1P20 RR16457 and American Cancer Society Grant RSG-04-247-01-CDD (to G. S.). 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.

^** Supported by the Heme training grant.

^¶ Supported by National Institutes of Health Training Grant GM 07752.

To whom correspondence should be addressed. Tel.: 858-822-3360; Fax: 858-822-3361; E-mail: joeadams{at}ucsd.edu.

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

This article has been cited by other articles:

				S. Na, O. Collin, F. Chowdhury, B. Tay, M. Ouyang, Y. Wang, and N. Wang Rapid signal transduction in living cells is a unique feature of mechanotransduction PNAS, May 6, 2008; 105(18): 6626 - 6631. [Abstract] [Full Text] [PDF]

				M. M. Keshwani and T. K. Harris Kinetic Mechanism of Fully Activated S6K1 Protein Kinase J. Biol. Chem., May 2, 2008; 283(18): 11972 - 11980. [Abstract] [Full Text] [PDF]

				M. Vielreicher, G. Harms, E. Butt, U. Walter, and A. Obergfell Dynamic Interaction between Src and C-terminal Src Kinase in Integrin {alpha}IIbbeta3-mediated Signaling to the Cytoskeleton J. Biol. Chem., November 16, 2007; 282(46): 33623 - 33631. [Abstract] [Full Text] [PDF]

				N. Dey, P. K. De, M. Wang, H. Zhang, E. A. Dobrota, K. A. Robertson, and D. L. Durden CSK Controls Retinoic Acid Receptor (RAR) Signaling: a RAR-c-SRC Signaling Axis Is Required for Neuritogenic Differentiation Mol. Cell. Biol., June 1, 2007; 27(11): 4179 - 4197. [Abstract] [Full Text] [PDF]

				M. Dinh, D. Grunberger, H. Ho, S. Y. Tsing, D. Shaw, S. Lee, J. Barnett, R. J. Hill, D. C. Swinney, and J. M. Bradshaw Activation Mechanism and Steady State Kinetics of Bruton's Tyrosine Kinase J. Biol. Chem., March 23, 2007; 282(12): 8768 - 8776. [Abstract] [Full Text] [PDF]

				S. A. Lieser, J. Shaffer, and J. A. Adams Src Tail Phosphorylation Is Limited by Structural Changes in the Regulatory Tyrosine Kinase Csk J. Biol. Chem., December 8, 2006; 281(49): 38004 - 38012. [Abstract] [Full Text] [PDF]

				K.-Y. Cheng, M. E. M. Noble, V. Skamnaki, N. R. Brown, E. D. Lowe, L. Kontogiannis, K. Shen, P. A. Cole, G. Siligardi, and L. N. Johnson The Role of the Phospho-CDK2/Cyclin A Recruitment Site in Substrate Recognition J. Biol. Chem., August 11, 2006; 281(32): 23167 - 23179. [Abstract] [Full Text] [PDF]

				X. Gao and T. K. Harris Steady-state Kinetic Mechanism of PDK1 J. Biol. Chem., August 4, 2006; 281(31): 21670 - 21681. [Abstract] [Full Text] [PDF]

				S. Lee, M. K. Ayrapetov, D. J. Kemble, K. Parang, and G. Sun Docking-based Substrate Recognition by the Catalytic Domain of a Protein Tyrosine Kinase, C-terminal Src Kinase (Csk) J. Biol. Chem., March 24, 2006; 281(12): 8183 - 8189. [Abstract] [Full Text] [PDF]