J. Exp. Med.
© The Rockefeller University Press
0022-1007/97/08/393/11 $2.00
Volume 186, Number 3, August 4, 1997 393-403

In Vitro Translation and Assembly of a Complete T Cell Receptor-CD3 Complex

By Johannes B. Huppa and Hidde L. Ploegh

From the Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

The T cell receptor for antigen (TCR) is a multisubunit complex that consists of at least seven polypeptides: the clonotypic, disulfide-linked / heterodimer that is noncovalently associated with the invariant polypeptides of the CD3 complex (CD3-, -, -) and , a disulfide-linked homodimer. We achieved the complete assembly of the human TCR in an in vitro transcription/translation system supplemented with dog pancreas microsomes by simultaneous translation of the messenger RNAs encoding the TCR-, - and CD3-, -, -, and - subunits. CD3-, one of the subunits that initiates the assembly of the TCR in living cells, forms misfolded, disulfide-linked homooligomers when translated alone. However, co-translation of one of its first binding partners in the course of assembly, CD3- or -, led to the expression of mainly monomeric and correctly folded  subunits, the only form we could detect as part of a properly assembled TCR complex. In the absence of these subunits, the ER-resident chaperone calnexin interacted with oligomeric, i.e. misfolded, structures of CD3- in a glycan-independent manner. A glycan-dependent interaction between CD3- and calnexin was mediated by CD3- and concerned only monomeric CD3- complexed with CD3-, but was dispensable for proper folding of CD3-. We suggest that in addition to its signaling function, CD3- serves as a monitor for proper subunit assembly of the TCR.

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