Structural and Functional Relationships of Laurate (omega-1)-Hydroxylase (P-450)
月桂酸 (omega-1)-羟化酶 (P-450) 的结构和功能关系
基本信息
- 批准号:63580153
- 负责人:
- 金额:$ 1.54万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for General Scientific Research (C)
- 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:
- 关键词:
项目摘要
cDNAs for chimeras of rabbit liver P-450s (laurate (omega-1)-hydroxylase and testosterone 16 alpha-hydrox- ylase) and site-specific mutants of the laurate hydroxylase were constructed, and expressed in yeast cells utilizing DNA manipulation techniques. The P-450s thus syntesized were purified, and their propenires were examined to identify domains of laurate (omega-1)-hydroxylase responsible for its substrate specificity.1. Two segments of laurate (omega-1)-hydroxylase covering residues 90-125 and 210-252 constitute the substrate binding domain and cooperate to fix the substrate on the hydroxylase molecule. The chimeras containing the sequences of the laurate hydroxylase in the both regions are active in the hydroxylation of the fatty acids, while the chimeras devoid of these sequences in either of the two regions were practically inactive and exhibited lower affinity for the fatty acids than the wild-type hydroxylase. The chimeras containing neither of these sequences could not bind t … More he substrate.2. When the the carboxy-terminal 28 residues of the laurate hydroxylase were replaced by the corresponding sequence of testosterone 16 alpha-hydroxylase, the laurate hydroxylase activity increased about 2.5 times and, moreover, a new stereospecific hydroxylase activity (16 beta-hydroxylation of testosterone) appeared. These observations suggest that the laurate hydroxylase contains a structure that is capable of binding testosterone at a proper orientation for the hydroxylation of the steroid at the beta position, but the carboxy-terminal segment of the laurate hydroxylase prevents the testosterone molecule from gaining access to the binding site while that of the testosterone hydroxylase does not.3. Replacement of Thr-301 and/or Thr-302 from the laurate hydroxylase by other amino acid residues affected the rates of the laurate and caprate hydroxylations at the omega-1 and the omega positions, indicating that these residues play an important role in recognizing the difference in the chain length between the two fatty acids and determining the position in the substrates to be hydroxylated. Less
构建兔肝P-450(月桂酸(ω-1)-羟化酶和睾酮16 α-羟化酶)嵌合体和月桂酸羟化酶位点特异性突变体的cDNA,并利用DNA操作技术在酵母细胞中表达。纯化如此合成的P-450,并检查它们的丙烯线以鉴定负责其底物特异性的月桂酸(ω-1)-羟化酶的结构域。覆盖残基90-125和210-252的月桂酸(ω-1)-羟化酶的两个片段构成底物结合结构域,并协同将底物固定在羟化酶分子上。在两个区域中含有月桂酸羟化酶序列的嵌合体在脂肪酸的羟基化中是活性的,而在两个区域中的任一个中缺乏这些序列的嵌合体实际上是无活性的,并且表现出比野生型羟化酶更低的对脂肪酸的亲和力。不含这两种序列的嵌合体不能结合T细胞。 ...更多信息 2.他是一个很好的人。当月桂酸羟化酶的羧基端28个残基被睾酮16 α-羟化酶的相应序列取代时,月桂酸羟化酶活性增加约2.5倍,并且出现了新的立体特异性羟化酶活性(睾酮的16 β-羟化)。这些观察结果表明,月桂酸羟化酶含有一个结构,能够结合睾酮在一个适当的方向上的类固醇在β位置的羟基化,但月桂酸羟化酶的羧基末端部分阻止睾酮分子获得的结合位点,而睾酮羟化酶的不. 3.由其他氨基酸残基取代的Thr-301和/或Thr-302从月桂酸羟化酶的影响率的月桂酸和癸酸羟基化在ω-1和ω的位置,表明这些残基起着重要的作用,在识别的两种脂肪酸之间的链长的差异,并确定在底物中的位置被羟基化。少
项目成果
期刊论文数量(23)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Yoshio Imai, Tomohide Uno, Masahiko Nakamura, and Hiroshi Yokota: "Structure-Function Relationships of Cytochrome P-450 Laurate" Drug Metab. Rev., 20-2-4, 467-478, 1989.
Yoshio Imai、Tomohide Uno、Masahiko Nakamura 和 Hiroshi Yokota:“细胞色素 P-450 月桂酸酯的结构与功能关系”药物代谢。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Yoshio Imai and Masahiko Nakamura: "The Importance of Threonine-301 from Cytochrome P-450 (Laurate (omega-1)-Hydroxylase and Testosterone 16 alpha-Hydroxylase) in Substrate Binding as Demonstrated by Site-Directed Mutagenesis" FEBS Lett., 234-2, 313-315,
Yoshio Imai 和 Masahiko Nakamura:“定点诱变证明了细胞色素 P-450(月桂酸 (omega-1)-羟化酶和睾酮 16 α-羟化酶)中苏氨酸 301 在底物结合中的重要性”FEBS Lett.,234
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Tomohide Uno: "Replacing the Carboxy-Terminal 28 Residues of Rabbit Liver P-450(Laurate(ω-1)-Hydroxylase)with those of P-450 Testosterone 16α-Hydroxylase Produces a New Stereospecific Hydroxylase Activity" Biochem.Biophys.Res.Commun.(1990)
Tomohide Uno:“用 P-450 睾酮 16α-羟化酶的羧基端残基替换兔肝 P-450(月桂酸(ω-1)-羟化酶)的羧基端 28 个残基可产生新的立体特异性羟化酶活性”Biochem.Biophys.Res。通讯(1990)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Yoshio Imai: "Protein Engineering in Studies on Metabolism of Toxic Substances" J. Toxicol. Sci, 13-suppl., 290-295, 1988.
Yoshio Imai:“有毒物质代谢研究中的蛋白质工程”J. Toxicol。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
IMAI Yoshio其他文献
IMAI Yoshio的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('IMAI Yoshio', 18)}}的其他基金
Russian intelligentsia and the International Co-operative Movement
俄罗斯知识分子与国际合作化运动
- 批准号:
07610043 - 财政年份:1995
- 资助金额:
$ 1.54万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
High Pressure Synthesis and Ordered Structure Formation of Condensation Polymers
缩聚物的高压合成和有序结构形成
- 批准号:
05453142 - 财政年份:1993
- 资助金额:
$ 1.54万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Basic Research for New Polycondensation Systems Using Transition Metal Catalyst
使用过渡金属催化剂的新型缩聚体系的基础研究
- 批准号:
01470109 - 财政年份:1989
- 资助金额:
$ 1.54万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)