Altered assembly of spectrin in red cell membranes in hereditary pyropoikilocytosis.
Altered assembly of spectrin in red cell membranes in hereditary pyropoikilocytosis.
复制标题
遗传性焦细胞增多症中红细胞膜中血影蛋白的组装发生改变。
DOI：
10.1182/blood.v57.1.130.bloodjournal571130
复制
发表时间：
1981
期刊：
影响因子：
20.3
通讯作者：
R. Castleberry
中科院分区：
文献类型：
--
作者：
J. Palek;S. C. Liu;P. Liu;J. Prchal;R. Castleberry
遗传性焦嗜红细胞增多症（HPP）的红细胞不稳定，比正常红细胞更容易受到体外热损伤。我们已经研究了HPP不稳定性是否由膜中光谱蛋白组装的改变引起。我们通过分子间二硫偶联的氧化交联方法确定了最近的膜蛋白邻居。我们随后用聚丙烯酰胺凝胶电泳（PAGE）分离交联配合物。将它们进行还原裂解，用PAGE在二维空间上鉴定交联蛋白。为了防止在孵育期间由于ATP和GSH水平的降低而引起的膜蛋白组织的变化。我们用腺嘌呤进行厌氧培养。肌苷。和葡萄糖。当在46'C（15毫米）或37'C （bb0 16小时）孵育。HPP红细胞碎裂并转化为变细胞和球细胞。随后HPP红细胞与二胺或它们的鬼与CuSO4，邻苯三酚氧化交联产生高摩尔重量（bbb10 × 10’）的蛋白复合物
Red cells from hereditary pyropoikilocytosis (HPP) are unstable and more susceptible than normal to thermal damage in vitro. We have investigated whether or not the HPP instability results from alterations in spectrin assembly in the membrane. We identified the nearest membrane protein neighbors employing oxidative crosslinking by intermolecular disulfide couplings. We subsequently separated the crosslinked complexes by polyacrylamide gel electrophoresis (PAGE). subjected them to reductive cleavage and identified the crosslinked proteins by PAGE in the 2nd dimension. To prevent changes in membrane protein organization due to decrease of ATP and GSH levels during incubation. we used anaerobic incubation with adenine. inosine. and glucose. When incubated at 46’C (15 mm) or 37’C (>16 hr). HPP red cells were fragmented and converted to poikilocytes and spherocytes. Subsequent oxidative crosslinking of HPP red cells with diamide or of their ghosts with CuSO4, o-phenantrolinc produced a high mol wt (> 1 x 1 0’) protein complex