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J. Biol. Chem., Vol. 282, Issue 22, 16126-16134, June 1, 2007

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The Membrane-proximal Portion of CD3 Associates with the Serine/Threonine Kinase GRK2^*

Laura M. DeFord-Watts, Jennifer A. Young, Lisa A. Pitcher¹, and Nicolai S. C. van Oers²

From the Department of Immunology and Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9093

The activation of protein kinases is one of the primary mechanisms whereby T cell receptors (TCR) propagate intracellular signals. To date, the majority of kinases known to be involved in the early stages of TCR signaling are protein-tyrosine kinases such as Lck, Fyn, and ZAP-70. Here we report a constitutive association between the TCR and a serine/threonine kinase, which was mediated through the membrane-proximal portion of CD3 . Mass spectrometry analysis of CD3 -associated proteins identified G protein-coupled receptor kinase 2 (GRK2) as a candidate Ser/Thr kinase. Transient transfection assays and Western blot analysis verified the ability of GRK2 to interact with the cytoplasmic domain of CD3 within a cell. These findings are consistent with recent reports demonstrating the ability of certain G protein-coupled receptors (GPCR) and G proteins to physically associate with the / TCR. Because GRK2 is primarily involved in arresting GPCR signals, its interaction with CD3 may provide a novel means whereby the TCR can negatively regulate signals generated through GPCRs.

Received for publication, October 5, 2006 , and in revised form, March 22, 2007.

^* This work was supported in part by NIAID Grant 5T32AI005284 from the National Institutes of Health (to L. M. D.). 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 Present address: Dept. of Medicine, Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA 02215.

² Supported by an NIAID grant from the National Institutes of Health. To whom correspondence should be addressed: NA7.201, Dept. of Immunology, the University of Texas Southwestern Medical Center, 6000 Harry Hines Blvd., Dallas, TX 75390-9093. Tel.: 214-648-1236; Fax: 214-648-7331; E-mail: nicolai.vanoers{at}utsouthwestern.edu.

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