Cloning and Characterization of Cell Adhesion Kinase , a Novel Protein-tyrosine Kinase of the Focal Adhesion Kinase Subfamily

Department of Biochemistry, Sapporo Medical University School of Medicine, Japan.
Journal of Biological Chemistry (Impact Factor: 4.57). 10/1995; 270(36):21206-19. DOI: 10.1074/jbc.270.36.21206
Source: PubMed


A second protein-tyrosine kinase (PTK) of the focal adhesion kinase (FAK) subfamily, cell adhesion kinase beta (CAK beta), was identified by cDNA cloning. The rat CAK beta is a 115.7-kDa PTK that contains N- and C-terminal domains of 418 and 330 amino acid residues besides the central kinase domain. The rat CAK beta has a homology with mouse FAK over their entire lengths except for the extreme N-terminal 88 residues and shares 45% overall sequence identity (60% identical in the catalytic domain), which indicates that CAK beta is a protein structurally related to but different from FAK. The CAK beta gene is less evenly expressed in a variety of rat organs than the FAK gene. Anti-CAK beta antibody immunoprecipitated a 113-kDa protein from rat brain, 3Y1 fibroblasts, and COS-7 cells transfected with CAK beta cDNA. The tyrosine-phosphorylated state of CAK beta was not reduced on trypsinization, nor enhanced in response to plating 3Y1 cells onto fibronectin. CAK beta localized to sites of cell-to-cell contact in COS-7 transfected with CAK beta cDNA, in which FAK was found at the bottom of the cells. Thus, CAK beta is a PTK possibly participating in the signal transduction regulated by cell-to-cell contacts.

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    • "Focal adhesion kinase (FAK) family proteins have been described to play a role in cell migration during different conditions and paradigms (Tomar and Schlaepfer, 2009). Proline-rich tyrosine kinase 2 (Pyk2) is a non-receptor, calcium-dependent protein-tyrosine kinase that belongs to the FAK family (Lev et al., 1995; Sasaki et al., 1995). Whereas FAK is expressed in all tissues, Pyk2 is highly enriched in the brain, especially the forebrain where it is regulated by neuronal activity and may be involved in synaptic plasticity (Girault et al., 1999; Huang et al., 2001; Siciliano et al., 1996). "
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    ABSTRACT: Stimulation by chemokines of integrin α4β1-dependent T lymphocyte adhesion is a crucial step for lymphocyte trafficking. The adaptor Vav1 is required for chemokine-activated T-cell adhesion mediated by α4β1. Conceivably, proteins associating with Vav1 could potentially modulate this adhesion. Correlating with activation by the chemokine CXCL12 of T lymphocyte attachment to α4β1 ligands, a transient stimulation in the association of Vav1 with SLP-76, Pyk2 and ADAP was observed. Using T-cells depleted for SLP-76, ADAP or Pyk2, or expressing Pyk2 kinase inactive forms, we show that SLP-76 and ADAP stimulate chemokine-activated, α4β1-mediated adhesion, whereas Pyk2 opposes T-cell attachment. While CXCL12-promoted generation of high-affinity α4β1 is independent of SLP-76, ADAP and Pyk2, the strength of α4β1-VCAM-1 interaction and cell spreading on VCAM-1 are targets of regulation by these three proteins. GTPase assays, expression of activated or dominant negative Rac1, or combined ADAP and Pyk2 silencing indicated that Rac1 activation by CXCL12 is a common mediator response in SLP-76-, ADAP- and Pyk2-regulated cell adhesion involving α4β1. Our data strongly suggest that chemokine-stimulated associations between Vav1, SLP-76 and ADAP facilitate Rac1 activation and α4β1-mediated adhesion, whereas Pyk2 opposes this adhesion by limiting Rac1 activation. © 2015 by The American Society for Cell Biology.
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