Simone S M
Boll Soc Ital Biol Sper. 1983 Dec 30;59(12):1791-5.
An easy and rapid preparation of cytochrome b from beef heart mitochondria is described. The method is based on four fractionation steps in the presence of bile acid and one step DE chromatography. The isolated cytochrome shows only partial reactivity with carbon monoxide; up to a level of 35 micrograms protein applied, it moves as a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, with an apparent molecular weight of 32000 +/- 1000. A ratio of 14.1 +/- 0.8 nmol heme b/mg protein has been found for isolated cytochrome, indicating that good part of heme b is lost during the isolation. The low number of steps required for the isolation could allow the identification of the stage(s) responsible for the loss of heme moiety and indicate preparation improvements.
本文描述了一种从牛心线粒体中轻松快速制备细胞色素b的方法。该方法基于在胆汁酸存在下的四个分级分离步骤和一步DE色谱法。分离出的细胞色素与一氧化碳仅表现出部分反应性；在应用高达35微克蛋白质的水平下，它在十二烷基硫酸钠存在的情况下在聚丙烯酰胺凝胶电泳中作为单一谱带移动，表观分子量为32000±1000。已发现分离出的细胞色素的血红素b与蛋白质的比例为14.1±0.8纳摩尔/毫克，这表明在分离过程中相当一部分血红素b丢失了。分离所需的步骤较少，这可以确定导致血红素部分丢失的阶段，并指明制备方法的改进方向。
