Waugh R E, Agre P
Department of Biophysics, University of Rochester School of Medicine and Dentistry, New York 14642.
J Clin Invest. 1988 Jan;81(1):133-41. doi: 10.1172/JCI113284.
Hereditary spherocytosis is a common hemolytic anemia associated with deficiencies in spectrin, the principal structural protein of the erythrocyte membrane-skeleton. We have examined 20 different individuals from 10 spherocytosis kindreds and 2 elliptocytosis kindreds to determine the effects of different levels of spectrin deficiency on the viscoelastic properties of the erythrocyte membrane. Micropipettes were used to perform single-cell micromechanical measurements of approximately 1,000 individual cells to determine the membrane elastic shear modulus, the apparent membrane bending stiffness, and whole cell recovery time constant for the different cell populations. The membrane viscosity was calculated by the product of the shear modulus and the recovery time constant. Results show correlation between the fractional reduction in shear modulus and the fractional reduction in spectrin content (determined by spectrin radioimmunoassay) and spectrin density (determined by the ratios of spectrin to band 3 on electrophoresis gels) suggesting that membrane shear elasticity is directly proportional to the surface density of spectrin on the membrane (P less than 0.001). The apparent membrane bending stiffness is also reduced in proportion to the density of spectrin (P less than 0.001). The membrane viscosity is reduced relative to control (P less than 0.001), but the nature of the relationship between spectrin density and membrane viscosity is less clearly defined. These studies document striking relationships between partial deficiencies of erythrocyte spectrin and specific viscoelastic properties of the mutant membranes.
遗传性球形红细胞增多症是一种常见的溶血性贫血,与红细胞膜骨架的主要结构蛋白血影蛋白缺乏有关。我们研究了来自10个球形红细胞增多症家族和2个椭圆形红细胞增多症家族的20个不同个体,以确定不同程度的血影蛋白缺乏对红细胞膜粘弹性特性的影响。使用微量移液器对大约1000个单个细胞进行单细胞微机械测量,以确定不同细胞群体的膜弹性剪切模量、表观膜弯曲刚度和全细胞恢复时间常数。膜粘度通过剪切模量与恢复时间常数的乘积计算得出。结果表明,剪切模量的分数降低与血影蛋白含量(通过血影蛋白放射免疫测定法确定)和血影蛋白密度(通过电泳凝胶上血影蛋白与带3的比率确定)的分数降低之间存在相关性,这表明膜剪切弹性与膜上血影蛋白的表面密度成正比(P小于0.001)。表观膜弯曲刚度也与血影蛋白密度成比例降低(P小于0.001)。膜粘度相对于对照降低(P小于0.001),但血影蛋白密度与膜粘度之间关系的性质不太明确。这些研究证明了红细胞血影蛋白部分缺乏与突变膜特定粘弹性特性之间的显著关系。