Matsumoto N, Shiomura T, Ishihara T, Adachi H, Miwa S
Tohoku J Exp Med. 1976 Dec;120(4):365-76. doi: 10.1620/tjem.120.365.
Human red cell membrane proteins were extracted by incubation of the ghost with hypotonic phosphate buffer (pH 7.4), N-ethylmaleimide and p-hydroxy-mercuribenzoate. In paroxysmal nocturnal hemoglobinuria (PNH), hereditary spherocytosis (HS) and hereditary elliptocytosis, the amount of proteins extracted by these procedures was significantly less than the amount extractable from the ghost of normal and aplastic anemia red cells. Polypeptide patterns of red cell membranes in these hematological disorders were essentially similar to those of normal ghosts. Analysis of the supernatant by SDS polyacrylamide gel electrophoresis revealed that this reduction was mainly due to the reduced amount of peripheral proteins extracted. The extraction of peripheral proteins by sulfhydryl reagents was accompanied by shape changes resulting in the formation of membrane vesicles, suggesting an important role of peripheral proteins in the maintenance of ghost shape. It is also suggested that qualitative abnormalities of peripheral proteins such as altered reactivity to sulfhydryl reagents and/or strong binding to the membrane are present in PNH, HS and hereditary elliptocytosis red cells.
通过将血影与低渗磷酸盐缓冲液（pH 7.4）、N-乙基马来酰亚胺和对羟基汞苯甲酸盐一起孵育来提取人红细胞膜蛋白。在阵发性夜间血红蛋白尿（PNH）、遗传性球形红细胞增多症（HS）和遗传性椭圆形红细胞增多症中，通过这些方法提取的蛋白量明显少于从正常和再生障碍性贫血红细胞血影中可提取的蛋白量。这些血液系统疾病中红细胞膜的多肽模式与正常血影基本相似。通过SDS聚丙烯酰胺凝胶电泳对上清液进行分析表明，这种减少主要是由于提取的外周蛋白量减少。巯基试剂对外周蛋白的提取伴随着形状变化，导致形成膜泡，这表明外周蛋白在维持血影形状中起重要作用。还表明PNH、HS和遗传性椭圆形红细胞增多症红细胞中存在外周蛋白的定性异常，如对巯基试剂反应性改变和/或与膜的强结合。
