MOLECULAR CHARACTERIZATION OF THE HUMAN RED-CELL RHO(D) ANTIGEN
MOLECULAR CHARACTERIZATION OF THE HUMAN RED-CELL RHO(D) ANTIGEN
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DOI：
10.1002/j.1460-2075.1983.tb01409.x
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发表时间：
1983-01-01
期刊：
影响因子：
11.4
通讯作者：
GAHMBERG, CG
中科院分区：
文献类型：
--
作者：
GAHMBERG, CG
采用乳过氧化物酶125 I法对Rh血型-D/-D-（super-D）、-/-（Rhnull）的人红细胞和正常Rho（D）+细胞进行放射性表面标记。在SDS [十二烷基硫酸钠]存在下进行聚丙烯酰胺凝胶电泳，然后进行荧光分析，结果显示125 I标记的-D-/-Dd膜中明显富集了表观分子量为28，000 - 33，000的多肽。该多肽用抗Rho（D）抗血清特异性免疫沉淀。用胰蛋白酶或链霉蛋白酶处理完整细胞不能消化蛋白质。Rho多肽在还原和非还原条件下在聚丙烯酰胺凝胶电泳上迁移相同。在体外孵育红细胞与32 P后，它没有磷酸化。当整个标记的膜溶解在中性洗涤剂中并施加到凝集素-琼脂糖柱时，Rho（D）多肽吸附蓖麻凝集素，但不吸附小麦胚芽凝集素或透镜culinaris凝集素。纯化后的分子不与R. communis lectin-Sepharose.用内切-N-乙酰氨基葡糖苷酶H、内切-β-葡糖苷酶H处理Rho（D）抗原半乳糖苷酶或弱碱不降低其表观分子量。
Human red cells of Rh blood groups -D/-D- (super-D), ---/--- (Rhnull) and normal Rho(D) + cells were radioactively surface-labeled using the lactoperoxidase 125I method. Polyacrylamide gel electrophoresis in the presence of SDS [sodium dodecyl sulfate] followed by fluorography showed a strong enrichment of a polypeptide with an apparent MW of 28,000-33,000 in the 125I-labeled-D-/-Dd membranes. This polypeptide was specifically immune precipitated with anti-Rho(D) antiserum. Treatment of intact cells with trypsin or Pronase did not digest the protein. The Rho polypeptide migrated identically on polyacrylamide gel electrophoresis under reducing and nonreducing conditions. It was not phosphorylated after in vitro incubation of red cells with 32P. When whole labeled membranes were solubilized in neutral detergent and applied to lectin-Sepharose columns the Rho(D) polypeptide adsorbed to Ricinus communis lectin but not to wheat germ lectin or Lens culinaris lectin. The purified molecule did not adsorb to R. communis lectin-Sepharose. Treatment of the Rho(D) antigen with endo-N-acetyl glucosaminidase H, endo-.beta.-galactosidase or mild alkali did not lower its apparent MW.