Polymorphism in the Mr 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes.
Polymorphism in the Mr 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes.
复制标题
从 Rh(D) 阳性和阴性红细胞中纯化的 Mr 32,000 Rh 蛋白的多态性。
DOI：
10.1073/pnas.85.11.4042
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发表时间：
1988
影响因子：
11.1
通讯作者：
Agre,P
中科院分区：
文献类型：
--
作者：
Saboori,AM;Smith,BL;Agre,P
先前已在携带Rh（D）抗原的红细胞上鉴定出Mr 32，000的整合膜蛋白，并认为其含有导致不同Rh表型的抗原变异。为了在生化水平上研究它，开发了一种简单的大规模方法来从多个单位的Rh（D）阳性和阴性血液中纯化Mr 32，000 Rh蛋白。将红细胞膜囊泡溶解在NaDodSO 4中，并加入免疫沉淀的125 I表面标记的Rh蛋白的示踪剂。Rh蛋白通过羟基磷灰石色谱法纯化至均一，然后通过制备型NaDodSO 4/PAGE。从每单位Rh（D）阳性和Rh（D）阴性血液中回收约25 nmol纯Rh蛋白。从两种Rh表型中纯化的Rh蛋白通过一维NaDodSO 4/PAGE显示相似，并且前20个残基的N-末端氨基酸序列是相同的。用~（125）I标记和α-糜蛋白酶消化后的二维碘肽图谱比较了从Rh（D）阳性和阴性血中纯化的Rh蛋白。肽图谱非常相似;然而，在从Rh（D）阳性红细胞纯化的Rh蛋白中始终观察到至少两种额外的碘肽。这些数据表明，Rh（D）阳性和Rh（D）阴性红细胞中存在相似的核心Rh蛋白（或一组相关蛋白），并且来自具有不同Rh表型的红细胞的Rh蛋白含有不同的结构多态性。
A Mr 32,000 integral membrane protein has previously been identified on erythrocytes bearing the Rh(D) antigen and is thought to contain the antigenic variations responsible for the different Rh phenotypes. To study it on a biochemical level, a simple large-scale method was developed to purify the Mr 32,000 Rh protein from multiple units of Rh(D)-positive and -negative blood. Erythrocyte membrane vesicles were solubilized in NaDodSO4, and a tracer of immunoprecipitated 125I surface-labeled Rh protein was added. The Rh protein was purified to homogeneity by hydroxylapatite chromatography followed by preparative NaDodSO4/PAGE. Approximately 25 nmol of pure Rh protein was recovered from each unit of Rh(D)-positive and -negative blood. Rh protein purified from both Rh phenotypes appeared similar by one-dimensional NaDodSO4/PAGE, and the N-terminal amino acid sequences for the first 20 residues were identical. Rh proteins purified from Rh(D)-positive and -negative blood were compared by two-dimensional iodopeptide mapping after 125I-labeling and alpha-chymotrypsin digestion. The peptide maps were very similar; however, at least two additional iodopeptides were consistently noted in the Rh proteins purified from Rh(D)-positive erythrocytes. These data indicate that a similar core Rh protein (or group of related proteins) exists in both Rh(D)-positive and -negative erythrocytes, and the Rh proteins from erythrocytes with different Rh phenotypes contain distinct structural polymorphisms.
获得性免疫性溶血性贫血
影响因子：
0.9
作者：
L. Petz;G. Garratty
通讯作者：
G. Garratty