Functional analysis of the T-cell-restricted protein tyrosine kinase Txk.
T lymphocytes express a range of tyrosine kinases that are involved in signalling processes driving cell activation, proliferation and differentation. Two tyrosine kinases expressed only in T cells, the Itk/Emt and Txk gene products, are members of the Tec family of kinases. The role of Tec kinases in cellular function is poorly understood, although a Tec kinase specific to B cells, Btk, is essential for B-cell development. To explore the contribution of the T-cell-specific Tec kinases to lymphocyte function, we have expressed human Txk in the baculovirus system and conducted the first characterization of its activity. We find that Txk exhibits a substrate preference in vitro quite distinct from that of the major T-cell kinases Lck and ZAP70, suggesting that Tec-family kinases might act on a distinct range of substrates. We also investigated the interactions of Txk with the cytoplasmic domains of the key signalling molecules CD3zeta, CD28 and CTLA4 and find that none of these are phosphorylated by Txk, nor are they ligands for the SH2 or SH3 domains of Txk. We conclude that it is unlikely that Txk has a role in the early signal transduction events associated with these key pathways controlling T-cell activation.
Immunopathology Unit, Glaxo Wellcome, Medicines Research Centre, Gunnels Wood Road, Stevenage, Herts. SG1 2NY, UK. jhe45655@glaxowellcome.co.uk
J H
JH
Ellis
R P
RP
Sutmuller
M J
MJ
Sims
S
S
Cooksley
D000069594:Abatacept; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics; D018106:CD28 Antigens / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D060908:CTLA-4 Antigen; D006801:Humans; D018796:Immunoconjugates; D007313:Insecta; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013379:Substrate Specificity; D013601:T-Lymphocytes / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism
D015703:Antigens, CD; D000943:Antigens, Differentiation; D018106:CD28 Antigens; D017252:CD3 Complex; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D018796:Immunoconjugates; D011994:Recombinant Proteins; D000069594:Abatacept; C089848:Tec protein-tyrosine kinase; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases