Heterodimeric, disulfide‐linked α/β T cell receptors in solution

Structural and functional analysis of T cell receptor (TcR)‐ligand binding would be greatly advanced by the availability of an intact, assembled TcR in soluble form. We have produced such a molecule, by splicing the extracellular domains of a TcR to the glycosyl phosphatidylinositol membrane anchor sequences of Thy‐1. The molecule is expressed in the absence of CD3 on the cell surface, and can be cleaved from the membrane by treatment with phosphatidylinositol‐specific phospholipase C. The α and β chains of the soluble molecule are paired in the native conformation as judged by reactivity with the anti‐Vβ8 monoclonal antibody F23.1, and with the anti‐clonotypic monoclonal antibody 1B2; it is a disulfide‐linked dimer with a mol. mass of 95 kDa on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis under nonreducing conditions, and 47 kDa after reduction. We conclude that we have generated an α/β TcR in soluble form.

[1]  N. Gascoigne.  Transport and secretion of truncated T cell receptor beta-chain occurs in the absence of association with CD3. , 1990, The Journal of biological chemistry.

[2]  D. Brown,et al.  Mechanism of membrane anchoring affects polarized expression of two proteins in MDCK cells. , 1989, Science.

[3]  C. Janeway,et al.  Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation. , 1989, Journal of immunology.

[4]  P. Marrack,et al.  Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. , 1989, Journal of immunology.

[5]  A. Weiss,et al.  The lysine residue in the membrane-spanning domain of the beta chain is necessary for cell surface expression of the T cell antigen receptor , 1988, The Journal of experimental medicine.

[6]  J. Lippincott-Schwartz,et al.  Degradation from the endoplasmic reticulum: Disposing of newly synthesized proteins , 1988, Cell.

[7]  B. Berkhout,et al.  Transfection of genes encoding the T cell receptor-associated CD3 complex into COS cells results in assembly of the macromolecular structure. , 1988, The Journal of biological chemistry.

[8]  J. Coligan,et al.  The biosynthesis and assembly of T cell receptor alpha- and beta-chains with the CD3 complex. , 1988, Journal of immunology.

[9]  B. Berkhout,et al.  Assembly of the human T cell receptor-CD3 complex takes place in the endoplasmic reticulum and involves intermediary complexes between the CD3-gamma.delta.epsilon core and single T cell receptor alpha or beta chains. , 1988, The Journal of biological chemistry.

[10]  V. Nussenzweig,et al.  Signal for attachment of a phospholipid membrane anchor in decay accelerating factor. , 1987, Science.

[11]  R. Flavell,et al.  Tissue-specific expression of cell-surface Qa-2 antigen from a transfected Q7b gene of C57BL/10 mice , 1987, The Journal of experimental medicine.

[12]  M. Davis,et al.  Secretion of a chimeric T-cell receptor-immunoglobulin protein. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Bluestone,et al.  Identification of a monoclonal antibody specific for a murine T3 polypeptide. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Bouvier,et al.  The major surface protein of Leishmania promastigotes is anchored in the membrane by a myristic acid‐labeled phospholipid. , 1986, The EMBO journal.

[15]  S. Tonegawa,et al.  Attachment of an anti-receptor antibody to non-target cells renders them susceptible to lysis by a clone of cytotoxic T lymphocytes. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[16]  P. Ralph.  Retention of lymphocyte characteristics by myelomas and theta + -lymphomas: sensitivity to cortisol and phytohemagglutinin. , 1973, Journal of immunology.

[17]  P. Magee,et al.  Effect of administration of the carcinogen dimethylnitrosamine on urinary 7-methylguanine. , 1967, The Biochemical journal.

[18]  Frank R Wilson.  N. R. A. , 1933 .

[19]  F. Oliveri,et al.  Solubilizing the T-cell receptor--problems in solution. , 1989, Immunology today.

[20]  P. Bjorkman,et al.  TCR recognition and selection in vivo. , 1989, Cold Spring Harbor symposia on quantitative biology.

[21]  R. Hyman,et al.  Somatic genetic analysis of the expression of cell surface molecules. , 1988, Trends in genetics : TIG.

[22]  H Clevers,et al.  The T cell receptor/CD3 complex: a dynamic protein ensemble. , 1988, Annual review of immunology.

[23]  D. Sherman,et al.  Immunoprecipitation of cell surface structures of cloned cytotoxic T lymphocytes by clone-specific antisera. , 1984, Proceedings of the National Academy of Sciences of the United States of America.