Raab M, Yamamoto M, Rudd C E
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
Mol Cell Biol. 1994 May;14(5):2862-70. doi: 10.1128/mcb.14.5.2862-2870.1994.
CD5 is a T-cell-specific antigen which binds to the B-cell antigen CD72 and acts as a coreceptor in the stimulation of T-cell growth. CD5 associates with the T-cell receptor zeta chain (TcR zeta)/CD3 complex and is rapidly phosphosphorylated on tyrosine residues as a result of TcR zeta/CD3 ligation. However, despite this, the mechanism by which CD5 generates intracellular signals is unclear. In this study, we demonstrate that CD5 is coupled to the protein-tyrosine kinase p56lck and can act as a substrate for p56lck. Coexpression of CD5 with p56lck in the baculovirus expression system resulted in the phosphorylation of CD5 on tyrosine residues. Further, anti-CD5 and anti-p56lck coprecipitated each other in a variety of detergents, including Nonidet P-40 and Triton X-100. Anti-CD5 also precipitated the kinase from various T cells irrespective of the expression of TcR zeta/CD3 or CD4. No binding between p59fyn(T) and CD5 was detected in T cells. The binding of p56lck to CD5 induced a 10- to 15-fold increase in p56lck catalytic activity, as measured by in vitro kinase analysis. In vivo labelling with 32P(i) also showed a four- to fivefold increase in Y-394 occupancy in p56lck when associated with CD5. The use of glutathione S-transferase-Lck fusion proteins in precipitation analysis showed that the SH2 domain of p56lck could recognize CD5 as expressed in the baculovirus expression system. CD5 interaction with p56lck represents a novel variant of a receptor-kinase complex in which receptor can also serve as substrate. The CD5-p56lck interaction is likely to play roles in T-cell signalling and T-B collaboration.
CD5是一种T细胞特异性抗原,它与B细胞抗原CD72结合,并在T细胞生长刺激过程中作为共受体发挥作用。CD5与T细胞受体ζ链(TcRζ)/CD3复合物相关联,并且由于TcRζ/CD3连接,其酪氨酸残基会迅速磷酸化。然而,尽管如此,CD5产生细胞内信号的机制尚不清楚。在本研究中,我们证明CD5与蛋白酪氨酸激酶p56lck偶联,并且可以作为p56lck的底物。在杆状病毒表达系统中,CD5与p56lck共表达导致CD5酪氨酸残基磷酸化。此外,抗CD5和抗p56lck在包括Nonidet P - 40和Triton X - 100在内的多种去污剂中相互共沉淀。抗CD5也能从各种T细胞中沉淀出该激酶,而与TcRζ/CD3或CD4的表达无关。在T细胞中未检测到p59fyn(T)与CD5之间的结合。通过体外激酶分析测量,p56lck与CD5的结合使p56lck催化活性增加了10至15倍。用32P(i)进行体内标记也显示,当与CD5结合时,p56lck中Y - 394位点的占有率增加了4至5倍。在沉淀分析中使用谷胱甘肽S - 转移酶 - Lck融合蛋白表明,p56lck的SH2结构域可以识别杆状病毒表达系统中表达的CD5。CD5与p56lck的相互作用代表了一种受体 - 激酶复合物的新型变体,其中受体也可以作为底物。CD5 - p56lck相互作用可能在T细胞信号传导和T - B协作中发挥作用。