Kiszkiss D F, Downey R J
J Bacteriol. 1972 Feb;109(2):803-10. doi: 10.1128/jb.109.2.803-810.1972.
Membranes were isolated from Bacillus stearothermophilus 2184D by lysozyme digestion of the cell wall and subsequent differential centrifugation. Observations with the electron microscope indicate that such membranes are relatively intact and have a typical membrane appearance. Nitrate will preferentially oxidize the cytochrome b of such membranes. Approximately 80% of the total respiratory nitrate reductase activity of whole cells can be localized in the washed membrane fraction and the process of membrane isolation results in a sixfold purification of this enzyme. Of several detergents tested, sodium dodecyl sulfate, Triton 114, and Triton X-100 are most effective in converting reduced methyl viologen-nitrate reductase to a form which will not pellet at 130,000 x g. Density gradient analysis reveals that such detergent-mediated solubilization converts virtually all membrane protein to a form of lighter density.
通过用溶菌酶消化细胞壁并随后进行差速离心，从嗜热脂肪芽孢杆菌2184D中分离出膜。电子显微镜观察表明，这种膜相对完整，具有典型的膜外观。硝酸盐将优先氧化这种膜中的细胞色素b。全细胞中约80%的总呼吸硝酸盐还原酶活性可定位于洗涤后的膜部分，并且膜分离过程使该酶纯化了六倍。在测试的几种去污剂中，十二烷基硫酸钠、Triton 114和Triton X-100在将还原型甲基紫精 - 硝酸盐还原酶转化为在130,000 x g下不会沉淀的形式方面最有效。密度梯度分析表明，这种去污剂介导的溶解几乎将所有膜蛋白转化为密度较轻的形式。
