Regulation of Sex-Specific Forms of Cytochrome P-450 : Cytochrome P-450 in Liver Microsomes of Hamsters

细胞色素 P-450 性别特异性形式的调节 : 仓鼠肝微粒体中的细胞色素 P-450

• 批准号: 63490001
• 负责人: KAMATAKI Tetsuya
• 金额: $ 3.39万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63490001/
• 关键词: Cytochrome P-450; Hamster; Liver microsomes; Drug Metabolizing enzyme; Sex difference; Purification; チトクロ-ムP-450; ハムスター; 肝ミクロゾーム; チトクロームP-450

Three forms of cytochrome P-450 were purified to electrophoretical homogeneity on SDS-PAGE. One of the forms of cytochrome P-450 (P-450-fHS1) purified from liver microsomes of female hamster was considered to be mainly present in a low spin state as judged by absolute spectrum of the oxidized form and was cross-reactive with anti-P450-male antibodies. Other forms of cytochrome P-450 Purified from liver microsomes of male and female hamsters according to the cross-reactivity with anti-P-450-male antibodies were tentatively termed P-450-mHS2 and P-450-fHS2, respectively. Chromatographic behavior of P-450-mHS2 on DEAE-5PW column and hydroxylapatite column were identical to those of P-450-fHS2. Judging from the absolute spectra, both cytochromes P-450 were shown to be mainly in low spin states. N-Terminal amino acid sequence of P-450-mHS2 was identical with that of P-450-fHS2 when the first 20 amino acid sequence was compared. In addition, N-terminal amino acid sequence of P-450-fHS2 was f … More ound to be 80% homologous to those of P-450-mHS2 and P-450-fHS2. Western blot analysis of liver microsomes of male hamsters with anti-P-450-fHS1 antibodies showed that P-450-fHS1 or cytochrome P-450 immunochemically related to P-450-fHS1 was also present in liver microsomes of male hamsters. However, the amounts of P-450-fHS1 was found to be much higher in female than male. On the contrary, the content of cytochrome P-450 cross-reactive with anti-P-450-mHS2 antibodies in female hamster microsomes was comparable to that in male hamster microsomes. The content of P-450-fHS1 in liver microsomes of female hamsters was decreased by gonadectomy of female hamsters or the administration of testosterone. Treatment of gonadectomized female hamsters with testosterone, but not with estradiol, resulted in further decrease in the amount of P-450-fHS1. On the other hand, the content of P-450-fHS1 or cytochrome P-450 immunochemically related to P-450-fHS1 in male hamster microsomes was virtually unaffected by gonadectomy or the treatment with estradiol whereas the content of the cytochrome P-450 was increased by the treatment of gonadectomized male hamsters with estradiol. Both P-450-fHS2 and P-450-fHS2 were active for 6beta-hydroxylation of progesterone but not for testosterone hydroxylation. P-450-fHS2 catalyzed N-demethylations of aminopyrine, benzphetamine and erythromycin, and O-deethylation of 7-ethoxycoumarin in a recontituted system. P-450-mHS2 catalyzed N-demethylation of erythromycin and O-deethylation 7-ethoxycoumarin but not N-demethylations of aminopyrine and benzphetamine. Less

将三种形式的细胞色素P-450纯化为SDS-PAGE上的电泳同质性。从雌性仓鼠的肝微粒体中纯化的细胞色素P-450（P-450-FHS1）的一种形式被认为主要存在于低自旋状态下，这是由氧化形式的绝对光谱所判断的，并与抗P450玛尔抗体交叉反应。根据与抗P-450-Male抗体的交叉反应，从雄性和雌性仓鼠的肝微粒体纯化的其他形式的细胞色素P-450分别称为租用称为P-450-MHS2和P-450-FHS2。 DEAE-5PW色谱柱和羟基磷灰石柱上P-450-MHS2的色谱行为与P-450-FHS2相同。从绝对光谱来看，两个细胞色素p-450主要在低自旋状态下。当比较前20个氨基酸序列时，P-450-MHS2的N末端氨基酸序列与P-450-FHS2相同。此外，P-450-FHS2的N末端氨基酸序列与P-450-MHS2和P-450-FHS2同源的80％同源。抗P-450-FHS1抗体的雄性仓鼠的肝微粒体的蛋白质印迹分析表明，在男性仓鼠的肝微粒体中，也存在与P-450-FHS1相关的P-450-FHS1或细胞色素P-450免疫化学与P-450-FHS1相关的。但是，发现女性的P-450-FHS1量比男性高得多。相反，雌性仓鼠微粒体中的细胞色素P-450交叉反应与抗P-450-MHS2抗体的含量与雄性仓鼠微粒体中的含量相当。雌性仓鼠的性腺切除术或给药可改善雌性仓鼠肝微粒体中P-450-FHS1的含量。用睾丸激素治疗雌激素，但不能用雌二醇治疗，导致P-450-FHS1的量进一步减少。另一方面，男性仓鼠微粒体中与P-450-FHS1相关的P-450-FHS1或细胞色素P-450的含量实际上不受性腺切除术或雌二醇治疗的影响，而通过治疗gonadectommited catrad的含量会增加细胞色素P-450的含量。 P-450-FHS2和P-450-FHS2均活跃，用于孕酮的6Beta-羟基化，但不能用于睾丸激素羟基化。 P-450-FHS2催化了氨基吡啶，苯丙胺和红霉素的N-甲基化，以及在再连续系统中7-乙氧基酸乳蛋白酶的O-二甲基化。 P-450-MHS2催化红霉素和O-二甲基化的N-甲基化7-乙氧基酸乳蛋白蛋白蛋白，而不是氨基氨基氨酸和苯甲酰胺的N-甲基化。较少的

项目成果

Masayuki Komori,Kanako Nishio,Hiroaki Ohi,Mitsukasu Kitada and Tetsuya Kamataki: "Molecular cloning and sequence analysis of cDNA containing the entire coding region for human fetal liver cytochrome Pー450." J.Biochem.105. 161-163 (1989)

Masayuki Komori、Kanako Nishio、Hiroaki Ohi、Mitsukasu Kitada 和 Tetsuya Kamataki：“含有人胎儿肝细胞色素 P-450 完整编码区的 cDNA 的分子克隆和序列分析，J.Biochem.161-163”。

Masayuki Komori, Kanako Nishio, Hiroaki Ohi, Mitsukazu Kitada and Tetsuya Kamataki: "Molecular cloning and sequence analysis of cDNA containing the entire coding region for human fetal liver cytochrome P-450." J. Biochem.105. 161-163 (1989)

Masayuki Komori、Kanako Nishio、Hiroaki Ohi、Mitsukazu Kitada 和 Tetsuya Kamataki：“含有人胎儿肝细胞色素 P-450 完整编码区的 cDNA 的分子克隆和序列分析。”

Kazuhide Ohta,Michiyo Motoya,Masayuki Komori,Toshiaki Miura,Mitsukazu Kitada and Tetsuya Kamataki: "Interspecies homology of cylochrome P-450:Purification and toxicological significance of a high spin form of cytochrome P-450(P-450-D2)from liver microsome

Kazuhide Ohta、Michiyo Motoya、Masayuki Komori、Toshiaki Miura、Mitsukazu Kitada 和 Tetsuya Kamataki：“细胞色素 P-450 的种间同源性：来自肝脏的细胞色素 P-450 (P-450-D2) 高自旋形式的纯化和毒理学意义

Hiroaki Ohi, Miyuki Iwasaki, Masayuki Komori, Toshiaki Miura, Mitsukazu Kitada, Shoryo Hayashi and Testuya Kamataki: "Effects of serum testosterone level with buserelin on the activities of drug and testosterone hydroxylases and on the content of a male-s

Hiroaki Ohi、Miyuki Iwasaki、Masayuki Komori、Toshiaki Miura、Mitsukazu Kitada、Shoryo Hayashi 和 Testuya Kamataki：“布舍瑞林对血清睾酮水平对药物和睾酮羟化酶活性以及男性睾酮含量的影响

Kazuhide Ohta,Michiyo Motoya,Masayuki Komori,Toshiaki Miura,MitsuKazu Kitada and Tetsuya Kamatahi: "Interspecies homology of cytochrome Pー450:Purification and tozicological significance of a high spin form of cytochrome Pー450 (Pー450ーD2)from liver microsom

Kazuhide Ohta、Michiyo Motoya、Masayuki Komori、Toshiaki Miura、MitsuKazu Kitada 和 Tetsuya Kamatahi：“细胞色素 P-450 的种间同源性：来自肝脏的细胞色素 P-450 (P-450-D2) 高自旋形式的纯化和毒理学意义微粒体