Function of a novel metal-binding motif having oxidized cysteine residues
具有氧化半胱氨酸残基的新型金属结合基序的功能
基本信息
- 批准号:12440191
- 负责人:
- 金额:$ 4.16万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Nitrile hydratases (NHase) are metalloenzymes containing a non-heme or a non-corrin cobalt active center and catalyze the hydration of various nitriles to the corresponding amides. We have shown that Fe-type NHase of Rhodococcus sp. N771 has a novel metal-binding motif containing two oxidized cysteine ligands, cysteine-sulfenic acid (Cys-SOH) and cysteine-sulfinic acid (Cys-SO2H). In the present research, we studied functions of the novel metal-binding motif in nitrile hydratase family.1.Function of the oxidized cysteine ligands-Isobutyronitrile (IBN) had been reported as a competitive inhibitor with a Ki value of 5 μM. We found that authentic IBN was hydrated normally and that the impurity present in commercially available IBN, 2-cyano-2-propyl hydroperoxide (Cpx) inhibited NHase activity strongly. Cpx specifically oxidized the Cys-SOH ligand in the metal-binding motif, to inactivate the enzyme irreversibly n-Butyric acid (BA) is known to a stabilizing agent of NHase and used for purification, storage and most experiments reported. We studied how BA stabilizes NHase. We showed that BA protected αCys114-SOH from aerobic oxidation to Cys-SO_2H. Both results results demonstrated that the Cys-SOH structure of αCys114 is essential for the catalytic activity.2.We revealed that the Cys-SO2H modification was conserved not only in Co-type NHase but also in thiocyanate hydrolase (SCNase) whose amino acid sequences were well conserved with NHases. We also have shown that SCNase is a novel member of Co-type NHase. We constructed the expression system of apo-SCNase in E..coli, and succeeded in its crystallization. X-ray crystal structure analysis of apo-SCNase is currently underway.
氰基水合酶(NHase)是一类含有非血红素或非科林钴活性中心的金属酶,催化各种氰化物水合生成相应的酰胺类化合物。我们已经证明了红球菌的Fe型氨基转移酶。N771有一个新的金属结合基序,含有两个氧化半胱氨酸配体,半胱氨酸-磺酸(Cys-SOH)和半胱氨酸-亚磺酸(Cys-SO2H)。在本研究中,我们研究了氰基水合酶家族中新的金属结合基序的功能。1.氧化半胱氨酸配体异丁腈的功能被报道为竞争性抑制物,其Ki值为5μM。我们发现,正品异丁腈正常水合,商品异丁腈中的杂质2-氰基-2-丙基过氧化氢(CPx)对氨酶活性有强烈的抑制作用。CPX特异地氧化金属结合基序中的Cys-SOH配体,以不可逆失活该酶。丁酸(BA)是NHase的稳定剂,被用于纯化、储存和大多数实验报道。我们研究了BA对NHase的稳定作用。结果表明,BA对αCys114-SOH的有氧氧化为Cys-SO2H有保护作用。这两个结果都证明了αCys114的半胱氨酸结构对催化活性是必不可少的。2.我们发现,半胱氨酸修饰不仅在Co型氨酶中保守,而且在硫氰酸盐水解酶中也是保守的,其氨基酸序列与氨酶非常保守。我们还证明了SCNase是Co型NHase的一个新成员。我们构建了apo-SCNase在大肠杆菌中的表达系统,并对其进行了结晶。脱氧核糖核酸酶的X射线晶体结构分析目前正在进行中。
项目成果
期刊论文数量(30)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Endo,Isao: "What evidences were elucidated about photoreactive nitrile hydratase?"Journal Molecular Catalysis B : Enzymatic. 10. 81-86 (2000)
Endo,Isao:“关于光反应性腈水合酶阐明了哪些证据?”《分子催化 B 杂志:酶学》。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Tsujimura, Masanari: "A novel inhibitor for Fe-type nitrile hydratase 2-cyano-2-propyl hydroperoxide"Journal of American Chemical Society. 125. 11532-11538 (2003)
Tsujimura,Masanari:“Fe型腈水合酶2-氰基-2-丙基氢过氧化物的新型抑制剂”美国化学会杂志。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Endo, Isao: "What evidences were elucidated about photo-reactive nitrile hydratase?"Journal Molecular Catalysis B : Enzymatic. Vol.10. 81-86 (2000)
Endo, Isao:“关于光反应性腈水合酶阐明了哪些证据?”《分子催化 B 杂志:酶学》。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Piersma, Sander R.: "Arginine 56 mutation in beta subunit of nitrile hydratase : importance of hydrogen bonding to the non-heme iron center"Journal of Bioinorganic Chemistry. Vol.80. 283-288 (2000)
Piersma, Sander R.:“腈水合酶 β 亚基中的精氨酸 56 突变:氢键对非血红素铁中心的重要性”生物无机化学杂志。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Endo, Isao: "Fe-type nitrile hydratase"Journal of Bioinorganic chemistry. Vol.83. 247-253 (2001)
Endo,Isao:“Fe型腈水合酶”生物无机化学杂志。
- 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 }}
ODAKA Masafumi其他文献
ODAKA Masafumi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('ODAKA Masafumi', 18)}}的其他基金
Elucidation of catalytic mechanisms as well as subunit-assembly of nitrile hydratase family enzymes by using advanced structural studies
通过先进的结构研究阐明腈水合酶家族酶的催化机制和亚基组装
- 批准号:
24350082 - 财政年份:2012
- 资助金额:
$ 4.16万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Structural and functional studies on Chloroethene sensor kinase and its application for bioremediation
氯乙烯传感器激酶的结构和功能研究及其在生物修复中的应用
- 批准号:
23651063 - 财政年份:2011
- 资助金额:
$ 4.16万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Elucidation of mechanisms for catalytic center maturation and catalysis of nitrile hydratase family enzyme
阐明催化中心成熟和腈水合酶家族酶催化的机制
- 批准号:
21350089 - 财政年份:2009
- 资助金额:
$ 4.16万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Molecular mechanism for biogenesis and function of the cobalt reactioncenter of thiocyanate hydrolase
硫氰酸水解酶钴反应中心生物发生和功能的分子机制
- 批准号:
19350080 - 财政年份:2007
- 资助金额:
$ 4.16万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
相似海外基金
Strained alkenes as chemical probes for cysteine sulfenic acid
应变烯烃作为半胱氨酸磺酸的化学探针
- 批准号:
DE150101863 - 财政年份:2015
- 资助金额:
$ 4.16万 - 项目类别:
Discovery Early Career Researcher Award
Role of Cysteine Sulfenic Acid Formation in Compartmentalization of VEGF Signalin
半胱氨酸磺酸形成在 VEGF 信号蛋白区室化中的作用
- 批准号:
8445715 - 财政年份:2013
- 资助金额:
$ 4.16万 - 项目类别:
Role of Cysteine Sulfenic Acid Formation in Compartmentalization of VEGF Signalin
半胱氨酸磺酸形成在 VEGF 信号蛋白区室化中的作用
- 批准号:
8620710 - 财政年份:2013
- 资助金额:
$ 4.16万 - 项目类别:
Site-Specific Analysis of Human Cysteine Sulfenic Acid Protein Modifications
人半胱氨酸磺酸蛋白修饰的位点特异性分析
- 批准号:
7347362 - 财政年份:2007
- 资助金额:
$ 4.16万 - 项目类别:














{{item.name}}会员




