Ding A H, Nathan C F
Beatrice and Samuel A. Seaver Laboratory, Department of Medicine, Cornell University Medical College, New York, New York 10021.
Anal Biochem. 1988 Nov 15;175(1):22-9. doi: 10.1016/0003-2697(88)90355-7.
A cyanide-insensitive b-type cytochrome (cytochrome b559) is involved in the respiratory burst of phagocytes. Spectroscopic measurement of its content in polymorphonuclear leukocytes or macrophages is complicated by signals arising from contaminating Hb or mitochondrial cytochromes upon reduction by dithionite. These interferences can be overcome by reducing the sample with dithionite, aerating it, and reducing it a second time with dithionite, in the presence of 10 microM cyanide. Cyanide suppresses the response of the interfering proteins to the second dithionite reduction. This modified method allows an accurate determination of the content of cytochrome b559 using smaller numbers of cells than previously feasible.
一种对氰化物不敏感的b型细胞色素（细胞色素b559）参与吞噬细胞的呼吸爆发。在用连二亚硫酸盐还原时，由于污染的血红蛋白或线粒体细胞色素产生的信号，使得对多形核白细胞或巨噬细胞中其含量进行光谱测量变得复杂。在10微摩尔氰化物存在的情况下，通过用连二亚硫酸盐还原样品、对其进行曝气处理，然后再用连二亚硫酸盐进行第二次还原，可以克服这些干扰。氰化物会抑制干扰蛋白对第二次连二亚硫酸盐还原的反应。这种改进的方法允许使用比以前更少的细胞数量来准确测定细胞色素b559的含量。
