PHYSICAL ASSOCIATION OF THE CYTOPLASMIC DOMAIN OF CD2 WITH THE TYROSINE KINASES P56(LCK) AND P59(FYN)

In T lymphocytes, CD2 forms part of a loosely associated membrane comp lex which includes the T cell receptor (TcR) for antigen, the CD3 subu nits, CD4 or CD8, CD5 and the protein tyrosine kinases p56lck and p59f yn. The interaction of CD2 with tyrosine kinases in this complex provi des a possible mechanism for transmembrane signal transduction by CD2. We have investigated whether the interaction of CD2 with the kinases is dependent on other known members of the complex, or whether an inde pendent association can be observed. Using in vitro kinase assays with immune complexes precipitated from cell lysates, we demonstrate that CD2 can associate with p56lck and p59fyn in a rat thymoma line that do es not express CD4 or CD8, and in a TcR-negative Jurkat cell line. In TcR-positive Jurkat cells that express rat CD2, interaction of CD2 wit h p56lck and p59fyn was clearly seen, but it was absent in cells where the cytoplasmic tail of CD2 is truncated, indicating that the interac tions are mediated by the cytoplasmic region of CD2. Furthermore, usin g cells expressing CD2 molecules with partial truncations in the cytop lasmic domain, we show that the association of CD2 with p56lck is prog ressively lost as the cytoplasmic domain is shortened., and that the c apacity of the mutants to associate with p56lck correlates with their capacity to transduce transmembrane signals.