Madyastha K M, Krishnamachary N
Biochem Biophys Res Commun. 1986 Apr 29;136(2):570-6. doi: 10.1016/0006-291x(86)90478-x.
Microsomal b-type hemoprotein designated, cytochrome b555 of C.roseus seedlings was solubilized using detergents and purified by a combination of ion exchange chromatography and gel filtration to a specific content of 18.5 nmol per mg of protein. The purified cytochrome b555 was homogeneous and estimated to have an apparent molecular weight of 16500 on SDS-PAGE. The absorption spectrum of the reduced form has major peaks at 424, 525 and 555 nm. The alpha-band of the reduced form is asymmetric with a pronounced shoulder at 559 nm. The spectrum of the pyridine ferrohemochrome shows absorption peaks at 557, 524 and 418 nm indicating that the cytochrome has protoheme prosthetic group. The purified cytochrome is autoxidizable and does not combine with carbon monoxide, azide or cyanide. It is reducible by NADH in the presence of NADH-cytochrome b555 reductase partially purified from C.roseus microsomes.
使用去污剂溶解了玫红长春花幼苗中名为细胞色素b555的微粒体b型血红素蛋白，并通过离子交换色谱和凝胶过滤相结合的方法进行纯化，使其达到每毫克蛋白质18.5纳摩尔的特定含量。纯化后的细胞色素b555是均一的，在SDS-PAGE上估计其表观分子量为16500。还原形式的吸收光谱在424、525和555纳米处有主要峰值。还原形式的α带不对称，在559纳米处有明显的肩峰。吡啶亚铁血红素的光谱在557、524和418纳米处有吸收峰，表明该细胞色素具有原血红素辅基。纯化后的细胞色素可自动氧化，不与一氧化碳、叠氮化物或氰化物结合。在存在从玫红长春花微粒体中部分纯化的NADH-细胞色素b555还原酶的情况下，它可被NADH还原。
