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*
Department of Biology and Molecular Biology Institute, San Diego State University, San Diego, CA 92182;
Division of Allergy, La Jolla Institute for Allergy and Immunology, San Diego, CA 92121; and
Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037
Expressed in mast and T-cells/inducible T cell tyrosine kinase
(Emt/Itk) is a protein tyrosine kinase required for T cell Ag receptor
(TCR)-induced activation and development. A physical interaction
between Emt/Itk and TCR has not been described previously. Here, we
have utilized laser scanning confocal microscopy to demonstrate that
Ab-mediated engagement of the CD3
chain induces the membrane
colocalization of Emt/Itk with TCR/CD3. Removal of the Emt/Itk
pleckstrin homology domain (
PH-Emt/Itk) abrogates the association of
the kinase with the cell membrane, as well as its activation-induced
colocalization with the TCR complex and subsequent tyrosine
phosphorylation. The addition of a membrane localization sequence to
PH-Emt/Itk from Lck restores all of these deficiencies except the
activation-induced tyrosine phosphorylation. Our data suggest that the
PH domain of Emt/Itk can be replaced with another membrane localization
signal without affecting the membrane targeting and activation-induced
colocalization of the kinase with the TCR. However, the PH domain is
indispensable for the activation-induced tyrosine phosphorylation of
the kinase.
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