The extracellular domain of the zeta-chain is essential for TCR function

The zeta-chain homodimer is a key component in the TCR complex and exerts its function through its cytoplasmic immunoreceptor-tyrosine activation motif (1). The zeta-chain extracellular (EC) domain is highly conserved; however, its functional and structural contributions to the TCR signaling have not been elucidated. We show that the EC domain of the zeta homodimer is essential for TCR surface expression. To gain a more detailed structural and functional information about the zeta-chain EC domain, we applied a cysteine scanning mutagenesis to conserved amino acids of the short domain. The results showed that the interchain disulfide bridge can be displaced by seven or eight amino acids along the EC domain. The TCR signaling efficacy was dramatically reduced during peptide/MHC engagement in the zeta mutants containing the displaced disulfide bond. These signaling defective zeta mutants produced an unconventional early tyrosine phosphorylation pattern. While the tyrosine phosphorylated forms of zeta (p21 and p23) could be observed during Ag stimulation, downstream signaling events such as the generation of phospho-p36, higher m.w. forms of phospho-zeta, and phospho-zeta/ZAP-70 complexes were impaired. Together these results suggest an important function of the phylogenetically conserved zeta-EC domain.