Acuto O, Fabbi M, Smart J, Poole C B, Protentis J, Royer H D, Schlossman S F, Reinherz E L
Proc Natl Acad Sci U S A. 1984 Jun;81(12):3851-5. doi: 10.1073/pnas.81.12.3851.
To obtain information about the structural basis for T-cell antigen recognition, a T3-associated Ti receptor molecule was isolated from crude membranes of the REX human thymic tumor line and purified by affinity chromatography with an anti- clonotypic monoclonal antibody in conjunction with preparative gel electrophoresis. NH2-terminal amino acid sequencing of the beta subunit unambiguously identified the amino acids in positions 2-12. Comparative protein sequence analysis by computer search demonstrated that this Ti beta sequence bore weak, but definite, homology to the first framework of the variable region of human lambda light chain. Anti-sera to a synthetic peptide corresponding to positions 2-11 precipitated the denatured Ti beta subunit from REX, thus confirming the above sequence. This information suggests that the Ti beta subunit is distantly related to human immunoglobulin lambda light chain and, moreover, should be of use in the molecular cloning of the Ti beta gene.
为了获取有关T细胞抗原识别的结构基础的信息,从REX人胸腺肿瘤系的粗膜中分离出一种与T3相关的Ti受体分子,并通过用抗独特型单克隆抗体结合制备性凝胶电泳的亲和层析法进行纯化。对β亚基的氨基末端氨基酸测序明确鉴定出了第2至12位的氨基酸。通过计算机搜索进行的比较蛋白质序列分析表明,该Tiβ序列与人类λ轻链可变区的第一个构架具有微弱但确定的同源性。针对对应于第2至11位的合成肽的抗血清沉淀出了来自REX的变性Tiβ亚基,从而证实了上述序列。该信息表明,Tiβ亚基与人类免疫球蛋白λ轻链有远缘关系,而且,这对于Tiβ基因的分子克隆应是有用的。