Immunochemical analysis of the Kx protein from human red cells of different Kell phenotypes using antibodies raised against synthetic peptides
Immunochemical analysis of the Kx protein from human red cells of different Kell phenotypes using antibodies raised against synthetic peptides
复制标题
DOI：
10.1046/j.1365-2141.1997.d01-2110.x
复制
发表时间：
1997-03-01
影响因子：
6.5
通讯作者：
Bertrand, O
中科院分区：
文献类型：
--
作者：
Carbonnet, F;Hattab, C;Bertrand, O
Kx蛋白是一种在罕见的McLeod综合征患者中缺乏的红细胞膜多肽，最近克隆了该蛋白的编码基因，并独立纯化了Kx蛋白与Kell血型蛋白形成的共价复合体。为了进一步研究Kx膜蛋白，从该蛋白的一级序列衍生的6个合成肽在兔体内制备了抗血清，除2个抗血清外，其余抗血清均能在体外诱导体外转录-翻译合成的重组Kx蛋白。在纯化的Kell-Kx复合体和不同Kell血型变异体的SDS红细胞膜裂解物中，三种抗血清与免疫印迹反应，其中K-0缺少93kD的Kell蛋白。然而，没有发现与缺乏Kx蛋白的McLeod制剂的反应性，因此清楚地表明这些抗体具有Kx特异性。出乎意料的是，K-0细胞中Kx蛋白的相对含量低于具有共同Kell表型的红细胞，这表明Kell蛋白的缺失可能改变了膜上Kx的含量。
The Kx protein is an erythrocyte membrane polypeptide which is deficient in rare individuals suffering from the McLeod syndrome, The gene encoding this protein has been recently cloned and the Kx protein independently purified as a covalent complex with the Kell blood group protein, To further study the Kx membrane protein, antisera raised in rabbits against six synthetic peptides derived from the primary sequence of this protein were characterized, All antisera but two precipitated the recombinant Kx protein synthesized in coupled transcription-translation in vitro. Three antisera reacted on immunoblots with the 37 kD Kx protein present in the purified Kell-Kx complex and in SDS red cell membrane lysates from variants with different Kell blood group phenotypes, including K-0, which lack the Kell protein of 93 kD. However, no reactivity was found with McLeod preparations lacking Kx protein, thus clearly indicating that these antibodies have a Kx specificity. Unexpectedly, the relative amount of Kx protein in K-0 cells was found to be lower than in red cells with common Kell phenotypes, suggesting that the absence of the Kell protein may alter the amount of Kx in the membrane.