从牛多形核中性粒细胞中纯化细胞色素b558。

Purification of cytochrome b558 from bovine polymorphonuclear neutrophils.

作者信息

Morel F, Vignais P V

机构信息

Département de Recherche Fondamentale, Centre d'Etudes Nucléaires, Grenoble, France.

出版信息

Biochem Biophys Res Commun. 1987 Nov 30;149(1):46-55. doi: 10.1016/0006-291x(87)91603-2.

DOI:10.1016/0006-291x(87)91603-2

PMID:3689417

Abstract

A 110 fold purification of cytochrome b558 from resting bovine neutrophils has been achieved. The plasma membrane bound cytochrome b was extracted with aminoxide WS35, a non ionic detergent. The purification procedure included liquid column chromatography on CM-C50 Sephadex, chromatofocusing on the anion exchanger PBE94, and gel filtration on P30 Biogel. The purified preparation was characterized by an heme to protein (nmol/mg) ratio of 7.7. The isoelectric point of cytochrome b was at pH 6.5. Upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate three bands corresponding to apparent Mr 64,000, 56,000 and 20,000 were revealed by staining with Coomassie Blue.

摘要

已实现从静止的牛嗜中性粒细胞中对细胞色素b558进行110倍的纯化。用非离子去污剂氨氧化物WS35提取与质膜结合的细胞色素b。纯化过程包括在CM-C50葡聚糖凝胶上进行液相柱色谱、在阴离子交换剂PBE94上进行色谱聚焦以及在P30生物凝胶上进行凝胶过滤。纯化制剂的血红素与蛋白质（nmol/mg）比率为7.7。细胞色素b的等电点为pH 6.5。在十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳时，用考马斯亮蓝染色显示出三条带，其表观分子量分别为64,000、56,000和20,000。