Looking in New Directions for Origins and Cryptic Mechanisms of Enzyme Catalysis

寻找酶催化起源和神秘机制的新方向

• 批准号: 10636781
• 负责人: JUDITH P KLINMAN
• 金额: $ 69.02万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10636781
• 关键词: Active Sites; Address; Antibiotics; Area; Biochemical; Biological; Catalysis; Chemistry; Coupling; Development; Drug Targeting; Enzymes; Event; Family; Hand; Human; Hydrogen; Knowledge; Methodology; Motion; PQQ Cofactor; Pathway interactions; Peptides; Post-Translational Protein Processing; Property; Proteins; Reaction; Ribosomes; Vitamins; catalyst; cofactor; design; enzyme activity; enzyme mechanism; enzyme structure; novel; novel strategies; protein function; prototype; temporal measurement

Project Summary: This project is focused on understanding the physical and mechanistic properties of enzymes that underlie their exquisite function. In recent years, protein motions have been implicated as essential to achieve an extremely rapid catalysis of bond cleavage events at enzyme active sites. Methodology for the spatial and temporal resolution of such protein motions has been developed using enzyme prototypes that catalyze hydrogen and methyl transfer reactions. These studies are now being extended to the TIM barrel family of enzymes that represent 10% of known enzyme structures and catalyze 5 out of 7 known EC classes. With this knowledge in hand, new approaches arise for protein redesign, de novo design and drug targeting. A second emerging area in biological catalysis concerns the post-translational modification of peptides that have been synthesized at the ribosome. A combination of structural and biochemical probes is addressing the enigmatic pathway that produces the bacterial cofactor and vitamin, pyrroloquinoline quinone. As the result of a number of recent breakthrough observations, each of the catalysts within the pathway is now amenable for detailed mechanistic study. These enzymes have little or no precedence in humans, making the PQQ pathway a possible new target for antibiotic development.

项目摘要： 该项目的重点是理解酶的物理和机械特性 他们精致的功能。近年来，蛋白质运动被认为是实现 酶活性位点上键裂解事件的极快催化。空间和 使用催化的酶原型开发了这种蛋白质运动的时间分辨率 氢和甲基转移反应。这些研究现在已扩展到蒂姆·桶家族 代表已知酶结构的10％并催化7种已知EC类别的5％的酶。与此 掌握的知识，蛋白质重新设计，从头设计和药物靶向的新方法。第二 生物催化中的新兴区域涉及已肽的翻译后修饰 在核糖体上合成。结构和生化探针的结合正在解决这个神秘的问题 产生细菌辅助因子和维生素，吡咯烷酚喹酮的途径。由于数字的结果 在最近的突破性观察中，途径中的每个催化剂现在都可以详细介绍 机械研究。这些酶在人类中几乎没有或根本没有优先权，使PQQ途径A 抗生素开发的新目标。

项目成果

Temperature-dependent hydrogen deuterium exchange shows impact of analog binding on adenosine deaminase flexibility but not embedded thermal networks.

• DOI: 10.1016/j.jbc.2022.102350
• 发表时间: 2022-09
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Gao, Shuaihua;Zhang, Wenju;Barrow, Samuel L.;Iavarone, Anthony T.;Klinman, Judith P.
• 通讯作者: Klinman, Judith P.

Understanding Biological Hydrogen Transfer Through the Lens of Temperature Dependent Kinetic Isotope Effects.

• DOI: 10.1021/acs.accounts.8b00226
• 发表时间: 2018-09-18
• 期刊: Accounts of chemical research
• 影响因子: 18.3
• 作者: Klinman JP;Offenbacher AR
• 通讯作者: Offenbacher AR

Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway.

• DOI: 10.1016/bs.mie.2018.04.002
• 发表时间: 2018
• 期刊: Methods in enzymology
• 作者: Zhu W;Martins AM;Klinman JP
• 通讯作者: Klinman JP

Hydrogen-Deuterium Exchange within Adenosine Deaminase, a TIM Barrel Hydrolase, Identifies Networks for Thermal Activation of Catalysis.

• DOI: 10.1021/jacs.0c07866
• 发表时间: 2020-11-25
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Gao S;Thompson EJ;Barrow SL;Zhang W;Iavarone AT;Klinman JP
• 通讯作者: Klinman JP

Hydrogen-deuterium exchange reveals long-range dynamical allostery in soybean lipoxygenase.

氢-氘交换揭示了大豆脂氧合酶的长程动态变构。

• DOI: 10.1074/jbc.m117.817197
• 发表时间: 2018
• 期刊: The Journal of biological chemistry
• 作者: Offenbacher,AdamR;Iavarone,AnthonyT;Klinman,JudithP
• 通讯作者: Klinman,JudithP