Flynn T P, Johnson G J, Allen D W
Blood. 1981 Jan;57(1):59-65.
To investigate the membrane abnormalities that may play a pathophysiologic role in several hemolytic anemias we determined the density distribution on sucrose density gradients of human red blood cell (RBC) membranes from patients with these disorders, from normal controls, and from incubated normal RBC. We analyzed the fractions for membrane-adsorbed hemoglobin (Hb), globin, and nonglobin cytoplasmic proteins. The relationship between the cytoplasmic proteins adsorbed on the membranes and the specific gravity (SG) of the membranes was linear. An increase in SG of the entire membrane population was seen in Hb C disease due to adsorbed Hb. Subpopulations of membranes with increased SG due to adsorption of nonglobin protein were evident in the membranes from two splenectomized patients with hemolytic glucose-6-phosphate dehydrogenase (G6PD) variants. Dense membrane subpopulations found in RBC membranes from three splenectomized patients with Hb Köln were associated with adsorbed globin, while similar subpopulations in RBC membranes from three splenectomized patients with hereditary spherocytosis demonstrated increased SG due to adsorbed Hb. Splenectomized normals had no such abnormality in membrane density. Sucrose density gradients demonstrate that membrane bound cytoplasmic protein is characteristic of the RBC membranes in several hemolytic disorders. Additionally, gradients are useful for the isolation and further analysis of those subpopulations of RBC membranes with abnormal SG and exaggerated membrane protein abnormalities.
为了研究可能在几种溶血性贫血中发挥病理生理作用的膜异常，我们测定了患有这些疾病的患者、正常对照以及孵育后的正常红细胞（RBC）的人红细胞膜在蔗糖密度梯度上的密度分布。我们分析了各组分中的膜吸附血红蛋白（Hb）、珠蛋白和非珠蛋白细胞质蛋白。吸附在膜上的细胞质蛋白与膜的比重（SG）之间呈线性关系。由于吸附了Hb，在Hb C病中可见整个膜群体的SG增加。在两名脾切除的溶血性葡萄糖-6-磷酸脱氢酶（G6PD）变异患者的膜中，因非珠蛋白吸附导致SG增加的膜亚群很明显。在三名脾切除的Hb Köln患者的RBC膜中发现的致密膜亚群与吸附的珠蛋白有关，而在三名脾切除的遗传性球形红细胞增多症患者的RBC膜中类似的亚群因吸附Hb而SG增加。脾切除的正常人在膜密度方面没有这种异常。蔗糖密度梯度表明，膜结合细胞质蛋白是几种溶血性疾病中RBC膜的特征。此外，梯度对于分离和进一步分析那些SG异常且膜蛋白异常夸大的RBC膜亚群很有用。
