Gascoigne N R, Goodnow C C, Dudzik K I, Oi V T, Davis M M
Proc Natl Acad Sci U S A. 1987 May;84(9):2936-40. doi: 10.1073/pnas.84.9.2936.
To produce sufficient quantities of soluble T-cell receptor protein for detailed biochemical and biophysical analyses we have explored the use of immunoglobulin--T-cell receptor gene fusions. In this report we describe a chimeric gene construct containing a T-cell receptor alpha-chain variable (V) domain and the constant (C) region coding sequences of an immunoglobulin gamma 2a molecule. Cells transfected with the chimeric gene synthesize a stable protein product that expresses immunoglobulin and T-cell receptor antigenic determinants as well as protein A binding sites. We show that the determinant recognized by the anticlonotypic antibody A2B4.2 resides on the V alpha domain of the T-cell receptor. The chimeric protein associates with a normal lambda light chain to form an apparently normal tetrameric (H2L2, where H = heavy and L = light) immunoglobulin molecule that is secreted. Also of potential significance is the fact that a T-cell receptor V beta gene in the same construct is neither assembled nor secreted with the lambda light chain, and when expressed with a C kappa region it does not assemble with the chimeric V alpha C gamma 2a protein mentioned above. This indicates that not all T-cell receptor V regions are similar enough to immunoglobulin V regions for them to be completely interchangeable.
为了产生足够量的可溶性T细胞受体蛋白用于详细的生化和生物物理分析，我们探索了免疫球蛋白-T细胞受体基因融合的用途。在本报告中，我们描述了一种嵌合基因构建体，其包含T细胞受体α链可变（V）结构域和免疫球蛋白γ2a分子的恒定（C）区编码序列。用该嵌合基因转染的细胞合成一种稳定的蛋白质产物，该产物表达免疫球蛋白和T细胞受体抗原决定簇以及蛋白A结合位点。我们表明，抗独特型抗体A2B4.2识别的决定簇位于T细胞受体的Vα结构域上。嵌合蛋白与正常的λ轻链结合形成一种明显正常的四聚体（H2L2，其中H =重链，L =轻链）免疫球蛋白分子并被分泌。同样具有潜在意义的是，同一构建体中的T细胞受体Vβ基因既不与λ轻链组装也不分泌，并且当与Cκ区一起表达时，它不与上述嵌合VαCγ2a蛋白组装。这表明并非所有T细胞受体V区都与免疫球蛋白V区相似到足以完全互换。
