Purification and partial characterization of the erythrocyte Kx protein deficient in McLeod patients.
Purification and partial characterization of the erythrocyte Kx protein deficient in McLeod patients.
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McLeod 患者红细胞 Kx 蛋白缺陷的纯化和部分表征。
DOI：
10.1111/j.1432-1033.1995.0931m.x
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
O. Bertrand
中科院分区：
文献类型：
--
作者：
S. Khamlichi;P. Bailly;D. Blanchard;D. Goossens;J. Cartron;O. Bertrand
用抗93 kDa Kell血型蛋白的单抗，从人红细胞中免疫纯化了一种37 kDa的蛋白。针对纯化的复合体产生的兔抗体与93-kDa和37-kDa的蛋白在Western印迹上反应，能够免疫沉淀来自大量表达Kx抗原的K0红细胞，但不能来自McLeod综合征患者的红细胞，McLeod综合征是一种X连锁疾病，缺乏Kx抗原。更多的研究表明，37 kDa的蛋白质没有糖基化，并允许建立22个第一个N末端氨基酸的序列。该序列与最近克隆的在McLeod患者中缺失或突变的XK基因的预测蛋白产物相同[Ho，M.，Chelly，J.，Carter，N.，Danek，A.，Crocker，P.&摩纳哥，A.P.(1994)Cell 77,869-880]。我们的发现提供了强有力的证据，表明37 kDa的红细胞膜蛋白与XK结构基因产生的Kx蛋白相同，并证明Kx和Kell蛋白是以二硫键(S)在红细胞表面以复合体形式表达的两个亚基。
A 37-kDa protein was immunopurified from human erythrocytes as a complex with a monoclonal antibody directed against the Kell blood group protein of 93 kDa. A rabbit antibody raised against the purified complex reacted on a Western blot with the 93-kDa and 37-kDa proteins and was able to immunoprecipitate the 37-kDa component from K0 erythrocytes which express large amount of the Kx antigen, but not from erythrocytes of patients suffering from McLeod syndrome, a X-linked disorder in which the Kx antigen is lacking. Additional studies have shown that the 37-kDa protein is not glycosylated, and permitted the sequence of the 22 first N-terminal amino acids to be established. This sequence was identical to the predicted protein product of the XK gene cloned recently, which is deleted or mutated in McLeod patients [Ho, M., Chelly, J., Carter, N., Danek, A., Crocker, P. & Monaco, A. P. (1994) Cell 77, 869-880]. Our findings provide strong evidence that the 37-kDa red cell membrane protein is identical to the Kx protein produced by the XK structural gene and demonstrate that Kx and Kell proteins are two subunits expressed as a complex hold by disulfide bond(s) at the red cell surface.