Expression of human T cell receptor-gamma delta structural forms.
The human TCR-gamma delta occurs in three biochemically distinct forms (forms 1, 2bc, and 2abc). A 40-kDa TCR gamma-chain is disulfide-linked to the TCR delta-chain in form 1, whereas 40-kDa or 55-kDa TCR-gamma polypeptides are noncovalently associated with the TCR delta-chain in forms 2bc and 2abc, respectively. Sequence analysis of TCR-gamma cDNA clones indicates that form 1 utilizes the C gamma 1 gene segment, whereas forms 2bc and 2abc appear to use allelic C gamma 2 gene segments containing either two copies (b and c) or three copies (a, b, and c) of the CII exon, respectively. We transfected TCR-gamma cDNA encoding form 1 or form 2abc into the MOLT-13 cell line that expresses form 2bc. The transfected TCR gamma-chains associate with the resident MOLT-13 TCR-delta, normally part of form 2bc, to yield CD3-associated TCR-gamma delta heterodimers identical to those seen on the donor cell lines (form 1 or 2abc). These transfection experiments show directly that, 1) when a single TCR-delta subunit is available, the presence or absence of disulfide linkage between TCR gamma- and TCR delta-chains is controlled by the TCR gamma-chain, and 2) the difference in the amount of N-linked carbohydrate attached to the transfected TCR-gamma proteins of form 2bc vs form 2abc is influenced by the presence or absence of CII exon copy "a" which appears to alter the secondary and/or tertiary structure of these TCR gamma-chain constant regions, thereby affecting the attachment of N-linked glycans. In contrast to the similar structure and usage of C beta 1 and C beta 2, TCR-gamma delta forms show striking differences in structure and are not equally represented in peripheral blood. Although the role of each form is unknown, it is possible that variable or joining-gene segment selection events or functional differences account for their unequal usage.
Band
H
H
Laboratory of Immunochemistry, Dana-Farber Cancer Institute, Boston, MA.
Hochstenbach
F
F
Parker
C M
CM
McLean
J
J
Krangel
M S
MS
Brenner
M B
MB
D002460:Cell Line; D004220:Disulfides; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000235:genetics / Q000302:isolation & purification; D015458:Leukemia, T-Cell / Q000235:genetics; D008970:Molecular Weight; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification; D011948:Receptors, Antigen, T-Cell / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000032:analysis* / Q000378:metabolism; D014162:Transfection
D004220:Disulfides; D007127:Immunoglobulin Constant Regions; D010455:Peptides; D011948:Receptors, Antigen, T-Cell