Greenwalt D E
Holland Laboratory, American Red Cross, Rockville, Maryland 20855.
Protein Expr Purif. 1993 Feb;4(1):72-5. doi: 10.1006/prep.1993.1011.
A simple one-step protocol for the purification of the integral membrane protein CD36 from milk-fat-globule membranes of bovine mammary epithelial cells is described. Nonionic detergent extracts of membrane were chromatographed on hydroxylapatite and pure CD36 was eluted with 1 M NaCl. Other proteins of the milk-fat-globule membrane were eluted after CD36 with phosphate buffer. Human platelet CD36 bound to hydroxylapatite only after neuraminidase treatment. CD36 is an extremely hydrophobic and highly glycosylated protein and previous purification procedures have required multiple steps. Chromatography of CD36 on hydroxylapatite provides a simple and quick method of purification which does not sacrifice yield.
本文描述了一种从牛乳腺上皮细胞的乳脂肪球膜中纯化整合膜蛋白CD36的简单一步法方案。膜的非离子去污剂提取物在羟基磷灰石上进行色谱分离,纯CD36用1M NaCl洗脱。乳脂肪球膜的其他蛋白质在CD36之后用磷酸盐缓冲液洗脱。人血小板CD36仅在神经氨酸酶处理后才与羟基磷灰石结合。CD36是一种极度疏水且高度糖基化的蛋白质,以前的纯化程序需要多个步骤。CD36在羟基磷灰石上的色谱分离提供了一种简单快速的纯化方法,且不会牺牲产量。