Beyers A D, Spruyt L L, Williams A F
Medical Research Council Cellular Immunology Unit, Sir William Dunn School of Pathology, University of Oxford, England.
Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2945-9. doi: 10.1073/pnas.89.7.2945.
The T-cell antigen receptor (TCR) complex is the key structure involved in signal transduction in T cells. To analyze associations between the TCR complex and other molecules, immunoprecipitations were carried out, followed by phosphorylation of molecules in vitro by tyrosine kinases associated with the precipitated molecules. This provided a sensitive assay for molecular complexes, and associations were demonstrated between the TCR complex and the surface antigens CD2, CD4, or CD8 and CD5 in normal rat T cells. The complexes were readily seen in immunoprecipitates from Brij 96 but not Nonidet P-40 detergent extracts. The multimolecular complexes are associated with the internal tyrosine kinases p56lck and p59fyn. The presence of p56lck associated with CD4 or CD8 was also examined in early thymocytes, natural killer cells, and macrophages. The kinase was present in all cases except that of normal macrophages.
T细胞抗原受体(TCR)复合体是T细胞信号转导所涉及的关键结构。为了分析TCR复合体与其他分子之间的关联,进行了免疫沉淀,随后通过与沉淀分子相关的酪氨酸激酶在体外对分子进行磷酸化。这为分子复合体提供了一种灵敏的检测方法,并且在正常大鼠T细胞中证明了TCR复合体与表面抗原CD2、CD4或CD8以及CD5之间存在关联。在Brij 96的免疫沉淀中很容易看到这些复合体,但在非离子去污剂NP - 40提取物中则不然。多分子复合体与内部酪氨酸激酶p56lck和p59fyn相关。还在早期胸腺细胞、自然杀伤细胞和巨噬细胞中检测了与CD4或CD8相关的p56lck的存在情况。除了正常巨噬细胞外,在所有情况下都存在这种激酶。