Luketich J D, Melner M H, Guengerich F P, Puett D
Biochem Biophys Res Commun. 1983 Mar 16;111(2):424-9. doi: 10.1016/0006-291x(83)90323-6.
The mitochondrial and microsomal cytochrome P-450 contents of C57B1/6 mouse testis have been measured using difference spectroscopy on stable enzyme preparations containing the ferrous-carbon monoxide complex. Results were obtained on control animals (52 +/- 3 days of age) and on animals injected subcutaneously with human choriogonadotropin (0.017 micrograms/g body weight 24 h prior to sacrifice). The high ratio of testicular mitochondrial cytochrome oxidase to P-450, which has previously precluded measurements of basal P-450 levels, was overcome by using N,N,N',N'-tetramethyl-p-phenylene diamine to bypass site II, in combination with antimycin A to prevent reverse electron flow. The basal levels of mitochondrial and microsomal P-450 in mouse testis were 37.9 +/- 3.5 and 28.9 +/- 1.6 pmol/mg protein, respectively. Following administration of a desensitizing dose of gonadotropin, the respective values were lowered to 19.9 +/- 1.4 and 19.6 +/- 2.1 pmol/mg protein in 24 h. This is the first report of a gonadotropin-mediated decrease in mitochondrial P-450 and thus demonstrates that desensitization leads to alterations in both microsomal and mitochondrial P-450 in mouse testis.
利用差示光谱法对含有亚铁 - 一氧化碳复合物的稳定酶制剂进行测定,从而得出了C57B1/6小鼠睾丸中的线粒体和微粒体细胞色素P - 450含量。对对照动物(52±3日龄)以及在处死前24小时皮下注射人绒毛膜促性腺激素(0.017微克/克体重)的动物进行了检测。此前,由于睾丸线粒体细胞色素氧化酶与P - 450的比例过高,使得基础P - 450水平的测量受到阻碍。通过使用N,N,N',N'-四甲基对苯二胺绕过位点II,并结合抗霉素A以防止反向电子流,克服了这一问题。小鼠睾丸中线粒体和微粒体P - 450的基础水平分别为37.9±3.5和28.9±1.6皮摩尔/毫克蛋白质。给予脱敏剂量的促性腺激素后,24小时内相应的值分别降至19.9±1.4和19.6±2.1皮摩尔/毫克蛋白质。这是关于促性腺激素介导的线粒体P - 450降低的首次报道,因此表明脱敏会导致小鼠睾丸中的微粒体和线粒体P - 450均发生改变。