TY - JOUR
T1 - Heterodimeric, disulfide‐linked α/β T cell receptors in solution
AU - Slanetz, Alfred E.
AU - Bothwell, Alfred L.M.
PY - 1991/1
Y1 - 1991/1
N2 - 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.
AB - 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.
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U2 - 10.1002/eji.1830210127
DO - 10.1002/eji.1830210127
M3 - Article
C2 - 1671358
AN - SCOPUS:0026034864
SN - 0014-2980
VL - 21
SP - 179
EP - 183
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 1
ER -