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J. Biol. Chem., Vol. 281, Issue 50, 38529-38534, December 15, 2006

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Tec Kinase Itk Forms Membrane Clusters Specifically in the Vicinity of Recruiting Receptors^*

Qian Qi, Nisebita Sahu^¶, and Avery August¹

From the Center for Molecular Immunology and Infectious Disease and Department of Veterinary and Biomedical Sciences, Immunology and Infectious Disease Graduate Program, ^¶Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802

The Tec family of tyrosine kinases transduces signals from antigen and other receptors in cells of the hematopoietic system. In particular, interleukin-2 inducible T cell kinase (Itk) plays an important role in modulating T cell development and activation. Itk is activated by receptors via a phosphatidylinositol 3-kinase-mediated pathway, which results in recruitment of Itk to the plasma membrane via its pleckstrin homology domain. We show here that membrane localization of Itk results in the formation of clusters of at least two molecules within 80 Å of each other, which is dependent on the integrity of its pleckstrin homology domain. By contrast, the proline-rich region within the Tec homology domain, SH3 or SH2 domains, or kinase activity were not required for this event. More importantly, these clusters of Itk molecules form in distinct regions of the plasma membrane as only receptors that recruit phosphatidylinositol 3-kinase reside in the same membrane vicinity as the recruited Itk. Our results indicate that Itk forms dimers in the membrane and that receptors that recruit Itk do so to specific membrane regions.

Received for publication, September 27, 2006 , and in revised form, October 20, 2006.

^* This work was supported by grants from the National Institutes of Health (Grants AI51626 and AI065566) and the American Heart Association (Grant 0330036N) (to A. A.). 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 To whom correspondence should be addressed: Center for Molecular Immunology and Infectious Disease, Dept. of Veterinary and Biomedical Science, 115 Henning Bldg., The Pennsylvania State University, University Park, PA 16802. Tel.: 814-863-3539; Fax: 814-863-6140, E-mail: axa45{at}psu.edu.

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