IMMUNOCHEMICAL EVIDENCE FOR INDUCTION OF THE ALCOHOL-OXIDIZING CYTOCHROME-P-450 OF RABBIT LIVER-MICROSOMES BY DIVERSE AGENTS - ETHANOL, IMIDAZOLE, TRICHLOROETHYLENE, ACETONE, PYRAZOLE, AND ISONIAZID
IMMUNOCHEMICAL EVIDENCE FOR INDUCTION OF THE ALCOHOL-OXIDIZING CYTOCHROME-P-450 OF RABBIT LIVER-MICROSOMES BY DIVERSE AGENTS - ETHANOL, IMIDAZOLE, TRICHLOROETHYLENE, ACETONE, PYRAZOLE, AND ISONIAZID
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DOI：
10.1073/pnas.82.12.4065
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
COON, MJ
中科院分区：
文献类型：
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作者：
KOOP, DR;CRUMP, BL;COON, MJ
兔肝微粒体细胞色素P-450同工酶3a，也称为P-450ALC，先前在本实验室从给药乙醇或咪唑的动物中分离出来，纯化的细胞色素在重组的系统中被证明具有催化乙醇和其他醇氧化的加氧酶功能。虽然以各种方式处理的动物肝微粒体表现出增加的酒精氧化活性，但没有证据表明这是由于酶诱导还是由于影响活性的其他因素。P-450同工酶3a的免疫化学定量现在通过使用NaDodSO4/PAGE[十二烷基硫酸钠/聚丙烯酰胺凝胶电泳]/印迹和点印迹技术对该细胞色素纯化抗体进行。在肝微粒体中，同工酶3a的特异含量由2- 4倍增加，以诱导作用的大小依次为：异烟肼、三氯乙烯、吡唑、乙醇、咪唑和丙酮。同工酶3a在对照动物中约占P-450总量的5%，在丙酮处理下增加到27%。同工酶3a依赖的丁醇氧化活性，由抗体对各种微粒体制剂的抑制作用决定，发现随着细胞色素含量的增加成比例地增加。
Isozyme 3a of rabbit liver microsomal cytochrome P-450, also termed P-450ALC, was previously isolated in this laboratory from animals administered ethanol or imidazole, and the purified cytochrome was shown to function in the reconstituted system as an oxygenase in catalyzing the oxidation of ethanol and other alcohols. Although liver microsomes from animals treated in various ways exhibit increased alcohol-oxidizing activity, evidence was not available as to whether this was due to enzyme induction or to other factors influencing the activity. Immunochemical quantitation of P-450 isozyme 3a was now achieved by use of purified antibody to this cytochrome in NaDodSO4/PAGE [sodium dodecyl sulfate/polyacrylamide gel electrophoresis]/blotting and dot-blotting techniques. The specific content of isozyme 3a in liver microsomes was found to be increased from 2- to > 4-fold by administration of the following agents, in increasing order of effectiveness as inducers: isoniazid, trichloroethylene, pyrazole, ethanol, imidazole and acetone. Isozyme 3a represents about 5% of the total P-450 in control animals and is increased to as high as 27% by acetone treatment. Isozyme 3a-dependent butanol-oxidation activity, determined by the inhibitory effect of antibody on the various microsomal preparations, was found to increase proportionally with increased content of this cytochrome.