Yoshida T, Lorence R M, Choc M G, Tarr G E, Findling K L, Fee J A
J Biol Chem. 1984 Jan 10;259(1):112-23.
We have developed a chemically defined, minimal growth medium for Thermus thermophilus which is suitable for nutritional studies, isotopic enrichment, and genetic manipulation of the organism. Reliable procedures are described for the large scale purification of cytochrome c552 from the periplasm and for cytochrome c555,549 and cytochrome c1 aa3 from the plasma membrane. In contrast to a previous report (Fee, J. A., Choc, M. G., Findling, K. L., Lorence, R., and Yoshida, T. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 147-151) which suggested a molecular weight near 200,000, the cytochrome c1aa3 complex was shown by protein and amino acid analyses to have Mr approximately 93,000. Sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis and reversed phase high performance liquid chromatography, combined with amino acid analyses, revealed the presence of only two proteins in a 1:1 ratio: C-protein has Mr approximately 33,000, binds heme C, and is thought to correspond to cytochrome c1. A-protein has Mr approximately 55,000 and is thought to bind the four redox components (2 heme A and 2 Cu) of cytochrome aa3.
我们已经为嗜热栖热菌开发了一种化学成分明确的基本生长培养基，该培养基适用于该生物体的营养研究、同位素富集和基因操作。文中描述了从周质中大规模纯化细胞色素c552以及从质膜中纯化细胞色素c555、549和细胞色素c1 aa3的可靠方法。与之前的一份报告（Fee, J. A., Choc, M. G., Findling, K. L., Lorence, R., and Yoshida, T. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 147 - 151）表明其分子量接近200,000不同，通过蛋白质和氨基酸分析显示细胞色素c1aa3复合物的Mr约为93,000。十二烷基硫酸钠 - 尿素 - 聚丙烯酰胺凝胶电泳和反相高效液相色谱，结合氨基酸分析，揭示仅存在两种比例为1:1的蛋白质：C蛋白的Mr约为33,000，结合血红素C，被认为对应于细胞色素c1。A蛋白的Mr约为55,000，被认为结合细胞色素aa3的四种氧化还原成分（2个血红素A和2个铜）。
