Purification of HLA-A2 antigen, fluorescent labeling of its intracellular region, and demonstration of an interaction between fluorescently labeled HLA-A2 antigen and lymphoblastoid cell cytoskeleton proteins in vitro.
Purification of HLA-A2 antigen, fluorescent labeling of its intracellular region, and demonstration of an interaction between fluorescently labeled HLA-A2 antigen and lymphoblastoid cell cytoskeleton proteins in vitro.
复制标题
纯化 HLA-A2 抗原，对其胞内区域进行荧光标记，并在体外演示荧光标记的 HLA-A2 抗原与淋巴母细胞细胞骨架蛋白之间的相互作用。
DOI：
10.1021/bi00522a042
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发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Veatch,WR
中科院分区：
文献类型：
--
作者：
Pober,JS;Guild,BC;Strominger,JL;Veatch,WR
Jordan S. Pober，* Braydon C.杰克·吉尔Strominger和William R.维奇摘要：用同种特异性单克隆抗体免疫亲和层析法从淋巴母细胞膜上纯化了洗涤剂溶解的HLA-A和HLA-B抗原（p44，12）。缺乏HLA重链的跨膜部分和细胞内部分的木瓜蛋白酶溶解的HLA-A和-B抗原（p34，12）先前已通过该技术纯化至均一性[帕勒姆，P.（1979年）。254，8709-8712]。相反，用帕勒姆的方法从总淋巴母细胞膜的去污剂提取物中分离的HLA-A和-B抗原的沉淀物总是含有大量的肌动蛋白（5- 50%，取决于制备方法）。肌动蛋白与去污剂溶解的HLA-A和-B抗原紧密结合，并且在不使蛋白质变性或蛋白水解的情况下不能从溶解的复合物中除去。然而，通过在去污剂提取HLA-A和-B抗原之前从膜上选择性提取细胞骨架蛋白，可以制备去污剂增溶的不含肌动蛋白的HLA-A和-B抗原。去污剂溶解的HLA-A和-B抗原含有一个容易还原的半胱氨酸残基，而pa-蛋白溶解的抗原则不含有，即半胱氨酸残基位于跨膜或细胞内部分
Jordan S. Pober,* Braydon C. Guild, Jack L. Strominger, and William R. Veatch abstract: Detergent-solubilized HLA-A and-B antigens (p44, 12) have been purified from lymphoblastoidcell mem-branes by means of allospecific monoclonal antibody immu-noaffinity chromatography. Papain-solubilized HLA-A and-B antigens (p34, 12) which lack the transmembrane and in-tracellular portions of the HLA heavy chain have previously been purified to homogeneity by this technique [Parham, P.(1979)/. Biol. Chem. 254, 8709-8712]. In contrast, prepa-rations of HLA-A and-B antigens isolated from detergent extracts of total lymphoblastoid cell membranes by the method of Parham invariably contain a substantial amount of actin (5-50%, depending on the preparation). The actin is tightly bound to the detergent-solubilized HLA-A and-B antigens and could not be removed from the solubilized complex without denaturation or proteolysis of the proteins. Detergent-solubilized HLA-A and-B antigens free of actin can be prepared, however, by selective extraction of cytoskeletal proteins from the membrane prior to detergent extraction of the HLA-A and-B antigens. Detergent-solubilized HLA-A and-B antigens contain an easily reduced cysteine residue (s) whereas pa-pain-solubilized antigensdo not; ie, the cysteine residue (s) is located within the transmembrane or intracellular portion