Bansal S K, Love J, Gurtoo H L
Biochem Biophys Res Commun. 1983 Nov 30;117(1):268-74. doi: 10.1016/0006-291x(83)91570-x.
The major form of cytochrome P-450 isolated and purified from the hepatic microsomes of phenobarbital pretreated rats by sequential chromatography on n-octylamino-Sepharose 4B and DEAE-cellulose columns was found to be homogeneous by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. However, this cytochrome P-450 was resolved into three bands by high pressure liquid chromatography on an Anpac ion-exchange column. High pressure liquid chromatography isolated forms had similar molecular weights of 55,000 with lambda max of the CO-reduced difference spectrum at 450 nm and were found to be in the low spin state. The results demonstrate the effectiveness of high pressure liquid chromatography in the resolution of cytochrome P-450s of similar molecular weights but different net charges.
通过在正辛基氨基琼脂糖4B和二乙氨基乙基纤维素柱上进行连续层析，从苯巴比妥预处理大鼠的肝微粒体中分离纯化得到的细胞色素P-450的主要形式，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析发现是均一的。然而，这种细胞色素P-450在Anpac离子交换柱上进行高压液相色谱分析时被分离成三条带。高压液相色谱分离得到的形式具有相似的分子量55,000，一氧化碳还原差光谱的最大吸收波长在450nm，并且发现处于低自旋状态。结果表明高压液相色谱在分离分子量相似但净电荷不同的细胞色素P-450方面是有效的。
