Anstee D J, Tanner M J
Biochem J. 1974 Mar;138(3):381-6. doi: 10.1042/bj1380381.
The distribution of protein and blood-group-antigen activity obtained after butanol extraction of erythrocyte ;ghosts' under various conditions is described. Butanol extraction under low-ionic strength conditions results in the recovery of membrane protein in high yield in the aqueous phase. Blood-group-A activity is found in both the aqueous and butanol phases, whereas blood-group-P activity is confined to the butanol phase and blood-group-I and blood-group-MN activity are restricted to the aqueous phase. Much lower yields of protein are obtained in the aqueous phase when high-ionic-strength conditions are used. An appreciable amount of material is precipitated at the interface. Under these conditions blood-group-P activity is found only in the butanol phase, blood group-A activity in the butanol phase and interface material and only blood-group-MN activity in the aqueous phase. In contrast with previous reports no correlation could be demonstrated between the secretor status of the donors and the presence of blood-group-A activity in the aqueous phase after butanol extraction under any of the extraction conditions used. By using butanol extraction under high-ionic-strength conditions it is possible to isolate the blood-group-MN-active sialoglycoprotein in high yield from erythrocyte ;ghosts' by a simple procedure.
本文描述了在不同条件下,用丁醇提取红细胞“血影”后所得蛋白质及血型抗原活性的分布情况。在低离子强度条件下进行丁醇提取,可使膜蛋白在水相中高产率回收。血型A活性在水相和丁醇相中均有发现,而血型P活性局限于丁醇相,血型I和血型MN活性则仅存在于水相。当采用高离子强度条件时,水相中蛋白质的产率要低得多。有相当数量的物质沉淀在界面处。在此条件下,血型P活性仅存在于丁醇相,血型A活性存在于丁醇相和界面物质中,而仅血型MN活性存在于水相中。与先前的报道不同,在所采用的任何提取条件下,均未发现供体的分泌状态与丁醇提取后水相中血型A活性的存在之间存在相关性。通过在高离子强度条件下进行丁醇提取,能够通过一个简单的程序从红细胞“血影”中高产率地分离出血型MN活性的唾液糖蛋白。