Characterization of hepatic and pulmonary cytochromes P-450 in 3-methylcholanthrene-treated hamsters
Characterization of hepatic and pulmonary cytochromes P-450 in 3-methylcholanthrene-treated hamsters
复制标题
3-甲基胆蒽处理仓鼠肝和肺细胞色素 P-450 的表征
作者：
Minro Watanabe;H. Fujii;I. Sagami;Miwa Tanno
用3-甲基胆蒽（MC）处理叙利亚金仓鼠的肝微粒体，将两种主要形式的肝细胞色素P-450（肝P-450 MCI和P-450 MCII）纯化约5倍。肝P-450 MCI和P-450 MCII的纯化制剂每mg蛋白分别含有9.6和8.3 nmol细胞色素P-450（P-450），并且基本上不含NADPH-细胞色素c（P-450）还原酶（fpT）、NADH-细胞色素b5还原酶和细胞色素b5。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）测得肝P-450 MCI和P-450 mMCII的分子量分别为56 000和53 500。此外，从MC处理的仓鼠的肺微粒体中纯化了主要形式的P-450（P-450 MC），并且每mg蛋白含有14.2nmol P-450，并且估计单体分子量为56000，表明在仓鼠中与肝P-450 MCI的分子量相似。从吸收光谱的氧化形式的肝P-450 MCI和P-450 MCII的高自旋和低自旋铁血红素蛋白，分别和肺P-450 MC是相似的肝P-450 MCII在其血红素蛋白自旋状态。然而，在肝脏P-450 MCI、P-450 MCII和肺P-450 MC之间的CO还原形式中未观察到差异，均显示446.5 nm Soret带。在含有fpT和二月桂酰磷脂酰胆碱（DLPC）的重组体系中，肺P-450 MC能有效地催化苯并[a]芘（BP）的羟化，其生成速率为11.4mol/min/mol P-450，而肝P-450 MCI和P-450 MCII均表现出较低的水平，即P-450 MCII的生成速率为11.4mol/min/mol P-450。例如，在一个实施例中，0.49分别为0.54。这些发现表明MC处理的仓鼠肺和肝之间BP羟基化活性存在明显的组织差异。
Two major forms of hepatic cytochrome P-450 (hepatic P-450MCI and P-450MCII) were purified approximately 5-fold from liver microsomes in Syrian golden hamsters treated with 3-methylcholanthrene (MC). The purified preparations of hepatic P-450MCI and P-450MCII contained 9.6 and 8.3 nmol cytochrome P-450 (P-450) per mg protein, respectively, and were essentially free from NADPH-cytochrome c (P-450) reductase (fpT), NADH-cytochrome b5 reductase and cytochrome b5. By sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weights of hepatic P-450MCI and P-450 mMCII were estimated to be 56 000 and 53 500. Further, a major form of pulmonary P-450 (P-450MC) were purified from lung microsomes of MC-treated hamster, and contained 14.2 nmol P-450 per mg protein, and estimated to be 56 000 in monomeric molecular weight, indicating the similar molecular weight to hepatic P-450MCI in the hamster. From the absorption spectra the oxidized forms of hepatic P-450MCI and P-450MCII were high- and low-spin ferric hemoproteins, respectively, and pulmonary P-450MC was similar to hepatic P-450MCII in their hemoprotein spin state. No difference, however, was observed in the CO-reduced forms among hepatic P-450MCI, P-450MCII and pulmonary P-450MC, all exhibiting 446.5 nm Soret bands. In a reconstituted system containing fpT and dilauroylphos-phatidylcholine (DLPC), pulmonary P-450MC efficiently catalyzed benzo[a]pyrene (BP) hydroxylation at a rate of 11.4 mol formed per min per mol P-450, but hepatic P-450MCI and P-450MCII both exhibited lower levels, e. g., 0.49 and 0.54, respectively. These findings indicated a clear tissue difference in the activity of BP hydroxylation between lung and liver in MC-treated hamsters.
影响因子：
2.9
作者：
GUENGERICH, FP;DANNAN, GA;KAMINSKY, LS
通讯作者：
KAMINSKY, LS
微粒体细胞色素 P-450 的纯化和表征。
DOI：
10.3109/00498258209038945
发表时间：
1982
期刊：
Xenobiotica; the fate of foreign compounds in biological systems
影响因子：
--
作者：
Guengerich,FP;Dannan,GA;Wright,ST;Martin,MV;Kaminsky,LS
通讯作者：
Kaminsky,LS