人红细胞膜带3蛋白的提纯与鉴定

提出了一种分离纯化人红细胞膜带3蛋白的不含血型糖蛋白制剂的改良方法:先后用0.89％NaCl、20mM pH8.0磷酸钠和0.05％TritonX-100处理膜除去膜骨骼蛋白类和血型糖蛋白,再用自行设计的凝胶制备电泳装置进一步纯化。冰冻干燥的制剂是均质的,得率为18.5±2.85％,它的分子量、氨基酸组成和紫外吸收光谱与文献报道基本相同。

The Purification and Identification of Band 3 Protein from Human Red Cell Membrane

An improved method of large-scale isolation and purification of Band 3 protein from human red cell membranes has been developed. Membranes were treated sequentially with isosmotic buffered saline solution (0.89% NaCl, 20 mM sodium phosphate, pH 8.0) and 0.05% Triton X-100 in 1:5 water-diluted buffered saline solution to remove their membrane skeletal proteins and glycophorines. The Band 3 protein remaining in the membrane preparation was purified further by polyacrylamide gel preparative electrophoresis with a big column(20cm glass column of 4.8 cm o.d.). Purified and lyophilized Band 3 protein appeared to be homogeneous when analyzed by gel electrophoresis in SDS.The yields in the overall preparation was 18.5 ± 2.85‰The analytic results of the amino acid composition and molecular weight of the Band 3 protein preparation are also reported in this paper.