title: Importance of the T cell receptor alpha-chain transmembrane distal region for assembly with cognate subunits.
abstract: Antigen recognition by alphabeta T lymphocytes is mediated via the multisubunit TCR complex consisting of invariant CD3gamma,delta,epsilon and zeta chains associated with clonotypic TCRalpha and beta molecules. Charged amino acids located centrally within the TCRalpha transmembrane region are necessary and sufficient for assembly with the CD3deltaepsilon heterodimer. Previously, we have shown that deletion of 6-12 amino acids from the carboxy terminus of the TCRalpha-chain dramatically abrogates surface TCR expression, suggesting that the distal portion of the TCRalpha transmembrane region contains information that regulates the assembly and/or intracellular transport of TCR complexes. We have examined in more detail the molecular basis for reduced TCR expression in T cells bearing truncated TCRalpha chains. We found that in contrast to wild-type (wt), variant TCRalpha proteins missing the last nine C-terminal amino acids did not associate with core CD3gamma,delta,epsilon chains and were not assembled into disulphide-linked alphabeta heterodimers. The stability of newly synthesised wt and variant TCRalpha molecules was similar, showing that the abrogated surface TCR expression was not a consequence of impaired protein survival. Nevertheless, truncated TCRalpha chains still assembled with the chaperon protein calnexin in the endoplasmic reticulum, indicating that the distal portion of the TCRalpha transmembrane region is not essential for calnexin interaction. These data document a role for the distal portion of the TCRalpha transmembrane region in the assembly of TCR complexes and provide a molecular basis for reduced TCR expression in cells bearing truncated TCRalpha chains.
authors.affiliation: Department of Microbiology & Immunology, School of Medicine, East Carolina University, Greenville, NC, USA.
authors.forename: J G
authors.initials: JG
authors.lastname: Shelton
authors.forename: S
authors.initials: S
authors.lastname: Gülland
authors.forename: K
authors.initials: K
authors.lastname: Nicolson
authors.forename: K P
authors.initials: KP
authors.lastname: Kearse
authors.forename: B T
authors.initials: BT
authors.lastname: Bäckström
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D004220:Disulfides / Q000737:chemistry; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006825:Hybridomas; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism; D021122:Protein Subunits; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000378:metabolism; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D002135:Calcium-Binding Proteins; D004220:Disulfides; D009844:Oligosaccharides; D021122:Protein Subunits; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D037281:Calnexin