Zhang Z Q, Fournier A, Tan Y H
J Biol Chem. 1986 Jun 15;261(17):8017-21.
Radioiodinated human beta-interferon-Ser 17 (Betaseron) was reversibly cross-linked to Daudi cells by dithiobis(succinimidylpropionate). The radioactive ligand was cross-linked to three macromolecules forming labeled complexes of apparent Mr values of 130,000, 220,000, and 320,000. Betaseron, human alpha-interferon, human interleukin 2 but not recombinant human gamma-interferon competed with the labeled ligand for binding to these putative receptor(s). Human leukocyte-produced gamma-interferon competed weakly with 125I-Betaseron for binding to Daudi cells. The Betaseron-receptor complex(es) was purified by passage through a wheat germ lectin column followed by chromatography on an anti-interferon immunosorbent column and semipreparative gel electrophoresis. The cross-linked ligand-receptor complex was shown to be highly purified by sodium dodecyl sulfate and acetic acid:urea:Triton X-100 polyacrylamide gel electrophoresis. It can be dissociated into the labeled Betaseron (Mr = 17,000) ligand and a receptor moiety which has an apparent molecular weight of 110,000. The chromatographic behavior of the ligand-receptor complex on wheat germ lectin column suggests that the receptor is a glycoprotein. The described procedure yielded about 1 microgram of Betaseron receptor from 10(10) Daudi cells, estimated to contain a maximum of about 15 micrograms of the receptor.
放射性碘化人β-干扰素-Ser 17（β-干扰素）通过二硫代双（琥珀酰亚胺丙酸酯）与Daudi细胞可逆交联。放射性配体与三种大分子交联，形成表观分子量分别为130,000、220,000和320,000的标记复合物。β-干扰素、人α-干扰素、人白细胞介素2，但重组人γ-干扰素不能与标记配体竞争结合这些假定的受体。人白细胞产生的γ-干扰素与125I-β-干扰素竞争结合Daudi细胞的能力较弱。β-干扰素-受体复合物通过小麦胚凝集素柱，然后在抗干扰素免疫吸附柱上进行色谱分离和半制备凝胶电泳进行纯化。通过十二烷基硫酸钠和乙酸：尿素：Triton X-100聚丙烯酰胺凝胶电泳显示交联的配体-受体复合物高度纯化。它可以解离成标记的β-干扰素（分子量=17,000）配体和一个表观分子量为110,000的受体部分。配体-受体复合物在小麦胚凝集素柱上的色谱行为表明该受体是一种糖蛋白。所述方法从10^10个Daudi细胞中产生约1微克β-干扰素受体，估计最多含有约15微克该受体。
