Immunoprecipitation of the solubilized membrane receptor for IgE of human cultured lymphoblastoid cells.
Immunoprecipitation of the solubilized membrane receptor for IgE of human cultured lymphoblastoid cells.
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对人培养的类淋巴母细胞的 IgE 溶解膜受体进行免疫沉淀。
DOI：
10.1111/j.1365-3083.1981.tb00129.x
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发表时间：
1981
影响因子：
3.7
通讯作者：
Spiegelberg,HL
中科院分区：
文献类型：
--
作者：
Fritsche,R;Meinke,GC;Spiegelberg,HL
使用山羊抗受体抗血清对培养的人类淋巴母细胞的 IgE (Fcε) 推定 Fc 受体进行了表征。抗血清是针对与 IgE-Sepharose-4B 免疫吸收柱结合的 Wil-2WT 细胞的 NP-40 溶解细胞成分制备的。抗血清特异性抑制 125 I 标记的 IgE 与 Fcε+阳性 RPMI-8866 淋巴母细胞的结合。 Fcε 阴性 Raji 和 Molt-4 细胞以及 IgE 和 IgG 吸收抗血清不会改变这种抑制活性。 Raji 和 Molt-4 细胞广泛吸收的抗血清沉淀了 125 I-乳过氧化物酶标记的 RPMI-8866 细胞裂解放射性的 7%，但 Raji 和 Molt 细胞仅沉淀了 0.5-1.8%。 SDS-PAGE 放射自显影分析显示了两种主要的标记肽，分别为 86,000 和 47,000 mol。重量。 RPMI-8866 的免疫沉淀物减少，但 Raji 或 Molt-4 细胞裂解物的免疫沉淀物不减少。通过 Sepharose-6B 凝胶过滤测定，与抗血清反应的溶解的放射性标记膜成分的大约分子量在 NP-40 中溶解为 250,000，在 NP-40-4murea 中溶解为 125,000。 250,000 和 125,000 mol.wt。材料由 86,000 和 47,000 条肽组成。数据表明，抗受体抗血清抑制 125I 标记的 IgE 与 Fcε 受体阳性淋巴母细胞的结合，并表明该受体可能由两个非共价连接的多肽组成，它们在 NP-40 和 NP-40-4murea 中保持相关。
The putative Fc receptor for IgE (Fcε) of cultured human lymphoblastoid cells was characterized by using a goat anti‐receptor antiserum. The antiserum was prepared against NP‐40‐solubilized cell components of Wil‐2WT cells which bound to an IgE‐Sepharose‐4B immunoabsorbent column. The antiserum specifically inhibited binding of125I‐labelled IgE to Fcε+positive RPMI‐8866 lymphoblastoid cells. Absorption of the antiserum with Fcε‐negative Raji and Molt‐4 cells and with IgE and IgG did not changes this inhibitory activity. Antiserum extensively absorbed with Raji and Molt‐4 cells precipated 7% of the radioactivity of lysis of125I‐lactoperoxidase‐labelled RPMI‐8866 cells but only 0.5–1.8% of that Raji and Molt cells. Autoradiography of SDS‐PAGE analysis demonstrated two major labeled peptides of 86,000 and 47,000 mol. wt. in reduced immunoprecipitates from RPMI‐8866 but not from Raji or Molt‐4 cell lysates. As determined by Sepharose‐6B gel filtration, the approximate molecular weight of the solubilized radiolabelled membrane component that reacted with the antiserum was 250,000 solubilized in NP‐40 and 125,000 in NP‐40‐4murea. Both the 250,000 and 125,000 mol.wt. material consisted of the 86,000 and 47,000 peptides. The data demonstrative that the anti‐receptor antiserum inhibited binding of125I‐labelled IgE to Fcε‐receptor‐positive lymphoblastoid cells and suggest that the receptor may consist of two non‐covalently linked polypeptides that remain associated in NP‐40 and NP‐40‐4murea.