Mechanism and Inhibition of Pyruvate Phosphate Dikinase

丙酮酸磷酸二激酶的作用机制及抑制作用

• 批准号: 6874406
• 负责人: DEBRA DUNAWAY-MARIANO
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6874406
• 关键词: Mycobacterium tuberculosis; X ray crystallography; active sites; adenosine triphosphate; chemical kinetics; chemical models; cofactor; computer simulation; conformation; diffusion; divalent metal; enzyme induction /repression; enzyme mechanism; enzyme substrate complex; intermolecular interaction; microorganism metabolism; nuclear magnetic resonance spectroscopy; phosphoenolpyruvate; phosphotransferases; pyruvates; site directed mutagenesis

DESCRIPTION (provided by applicant): This revised application requests funds to continue studies of separate site catalysis by the enzyme pyruvate phosphate dikinase (PPDK). PPDK catalyzes the interconversion of ATP, Pi, and pyruvate with AMP, PPi and PEP by using two separate active sites linked by a carrier histidine residue. The overall goal of the proposed studies is to determine the mechanism by which the carrier histidine is transferred between active sites through precisely oriented and timed domain-domain docking steps. These studies will provide a deeper understanding of the forces controlling transient protein-protein complex formation in signal transduction pathways and template directed biosynthetic pathways. Five specific aims are listed. These are (1) determine the X-ray crystal structure of PPDK conformer 2, (2) determine the role of domain linkers in facilitating successful domain-domain docking, (3) determine the role of interactions between domain-domain interface residues in facilitating correct docking orientation and in optimizing residence time, (4) distinguish between a through-solvent-domain-diffusion model and a sliding-domain model and (5) identify the role of the PPDK homologue in Mycobacterium tuberculosis. This last aim will be carried out for the purpose of discovering a novel metabolic pathway operating with in this pathogen, as well as a novel target for drug design.

描述（由申请人提供）：本修订后的申请要求提供资金 继续研究丙酮酸磷酸酶的单独位点催化作用 二激酶（PPDK）。 PPDK 催化 ATP、Pi 和丙酮酸的相互转化 通过使用由载体连接的两个单独的活性位点与 AMP、PPi 和 PEP 组氨酸残基。拟议研究的总体目标是确定 载体组氨酸在活性位点之间转移的机制 通过精确定向和定时的域-域对接步骤。这些研究 将提供对控制瞬态的力的更深入的理解 信号转导途径和模板中蛋白质-蛋白质复合物的形成 定向生物合成途径。列出了五个具体目标。这些是 (1) 确定PPDK构象异构体2的X射线晶体结构，（2）确定 域链接器在促进成功的域-域对接中的作用，(3) 确定域-域界面残基之间相互作用的作用 促进正确的对接方向并优化停留时间，(4) 区分穿过溶剂域扩散模型和 滑动域模型和 (5) 确定 PPDK 同源物在 结核分枝杆菌。最后一个目标将为此目的而实现 发现一种在这种病原体中起作用的新代谢途径，如 以及药物设计的新靶标。

项目成果

Separate site catalysis by pyruvate phosphate dikinase as revealed by deletion mutants.

缺失突变体揭示了丙酮酸磷酸二激酶的单独位点催化作用。

• DOI: 10.1021/bi00007a013
• 发表时间: 1995
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Xu,Y;McGuire,M;Dunaway-Mariano,D;Martin,BM
• 通讯作者: Martin,BM

Analysis of sequence homologies in plant and bacterial pyruvate phosphate dikinase, enzyme I of the bacterial phosphoenolpyruvate: sugar phosphotransferase system and other PEP-utilizing enzymes. Identification of potential catalytic and regulatory motifs

植物和细菌丙酮酸磷酸二激酶、细菌磷酸烯醇丙酮酸酶 I：糖磷酸转移酶系统和其他 PEP 利用酶的序列同源性分析。

• DOI: 10.1021/bi00500a006
• 发表时间: 1990
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Pocalyko,DJ;Carroll,LJ;Martin,BM;Babbitt,PC;Dunaway-Mariano,D
• 通讯作者: Dunaway-Mariano,D

Characterization of the covalent enzyme intermediates formed during pyruvate phosphate dikinase catalysis.

丙酮酸磷酸二激酶催化过程中形成的共价酶中间体的表征。

• DOI: 10.1021/bi00058a014
• 发表时间: 1993
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Thrall,SH;Mehl,AF;Carroll,LJ;Dunaway-Mariano,D
• 通讯作者: Dunaway-Mariano,D

Crystal structure of the petal death protein from carnation flower.

康乃馨花瓣死亡蛋白的晶体结构。

• DOI: 10.1021/bi051779y
• 发表时间: 2005
• 期刊: Biochemistry.
• 作者: Teplyakov,Alexey;Liu,Sijiu;Lu,Zhibing;Howard,Andrew;Dunaway-Mariano,Debra;Herzberg,Osnat
• 通讯作者: Herzberg,Osnat

Investigation of the role of the domain linkers in separate site catalysis by Clostridium symbiosum pyruvate phosphate dikinase.

研究结构域接头在共生梭菌丙酮酸磷酸二激酶的单独位点催化中的作用。

• DOI: 10.1021/bi0113061
• 发表时间: 2001
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Wei,M;Ye,D;Dunaway-Mariano,D
• 通讯作者: Dunaway-Mariano,D