Linkage between catalytic mechanism and conformational dynamics in (beta/alpha)8-barrel enzymes

(β/α)8-桶酶催化机制与构象动力学之间的联系

基本信息

项目摘要

The catalytic power of enzymes is linked with its conformational dynamics, but the molecular determinants of this linkage are largely unclear. Here, we address this problem by analyzing in parallel the kinetic mechanisms and the dynamics of loops near the active sites of enzymes. We will use the (beta/alpha)8-barrel proteins indoleglycerol phosphate synthase (IGPS) and the cyclase subunit (HisF) of imidazole glycerol phosphate synthase (ImGPS) from mesophilic and thermophilic microorganisms. For the thermophilic and the mesophilic enzymes, the rates of the individual catalytic steps, substrate and product binding and release, chemical transformation, will be determined by using pre-steady state kinetic measurements at various temperatures to obtain a detailed picture of the enzymatic mechanisms. The information deduced from these experiments will then be correlated with the dynamics of loop movements. To this end, the beta1alpha1-loop at the active site of mesophilic and thermophilic IGPS will be spin labeled, and changes in its dynamics in the course of catalysis will be followed by EPR. For mesophilic and thermophilic HisF, motions of the beta/alpha-loops near the active site will be analyzed by NMR relaxation dispersion experiments. Our results should allow us to gain novel insight into the molecular nature of the coupling between protein dynamics and the crossing of energy barriers in enzyme catalysis. They should also reveal whether and how homologous mesophilic and thermophilic (beta/alpha)8-barrel enzymes differ in their catalytic strategies. A better understanding of the relationship between dynamics and catalytic efficiency will also be valuable for enzyme engineering.
酶的催化能力与其构象动力学有关,但这种联系的分子决定因素在很大程度上还不清楚。在这里,我们通过并行分析酶活性部位附近的动力学机制和环的动力学来解决这个问题。我们将使用来自中温和高温微生物的(β/α)8桶蛋白质吲哚甘油磷酸合成酶(IGPS)和咪唑甘油磷酸合成酶(ImGPS)的环化酶亚单位(HISF)。对于嗜热酶和中温酶,将通过在不同温度下的预稳态动力学测量来确定各个催化步骤的速率、底物和产物的结合和释放以及化学转化,以获得详细的酶机理图。从这些实验中得出的信息将与环路运动的动力学相关联。为此,中温和高温IGPS活性中心的β1α1环将被自旋标记,其在催化过程中的动力学变化将被EPR跟踪。对于中温和高温HISF,活性中心附近的β/α环的运动将通过核磁共振弛豫色散实验来分析。我们的结果应该允许我们对蛋白质动力学和酶催化中跨越能量障碍之间的耦合的分子本质有新的见解。它们还应该揭示同源的中温和高温(β/α)8桶酶在催化策略上是否以及如何不同。更好地理解动力学和催化效率之间的关系对酶工程也是有价值的。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ 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 }}

Professor Dr. Reinhard Sterner其他文献

Professor Dr. Reinhard Sterner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('Professor Dr. Reinhard Sterner', 18)}}的其他基金

Allosteric communication and subunit interaction specificity in glutamine amidotransferases
谷氨酰胺酰胺转移酶的变构通讯和亚基相互作用特异性
  • 批准号:
    249556939
  • 财政年份:
    2014
  • 资助金额:
    --
  • 项目类别:
    Research Grants
Coordination of the SPP 1170
SPP 1170 的协调
  • 批准号:
    5431219
  • 财政年份:
    2004
  • 资助金额:
    --
  • 项目类别:
    Priority Programmes
Evolution of (beta-alpha)8-barrel enzymes
(β-α)8-桶酶的进化
  • 批准号:
    5356621
  • 财政年份:
    2001
  • 资助金额:
    --
  • 项目类别:
    Research Grants
Aktivierung und katalytischer Mechanismus von Glutamin-Amidotransferasen, untersucht an der Imidazolglycerinphosphat-Synthase aus Thermotoga maritima
海栖热袍菌咪唑甘油磷酸合酶谷氨酰胺酰胺转移酶的激活及催化机制研究
  • 批准号:
    5184154
  • 财政年份:
    1999
  • 资助金额:
    --
  • 项目类别:
    Research Grants
Design of allosteric light regulation in multi-enzyme complexes
多酶复合物变构光调节的设计
  • 批准号:
    323255115
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
    Research Grants
Experimental reconstruction of the molecular evolution of atrazine-degrading enzymes
阿特拉津降解酶分子进化的实验重建
  • 批准号:
    501122718
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
    Research Grants

相似海外基金

Characterizing the link between protein dynamics and catalytic function to improve the design of enzyme biocatalysts
表征蛋白质动力学和催化功能之间的联系,以改进酶生物催化剂的设计
  • 批准号:
    RGPIN-2022-04368
  • 财政年份:
    2022
  • 资助金额:
    --
  • 项目类别:
    Discovery Grants Program - Individual
Characterizing the link between protein dynamics and catalytic function to improve the design of enzyme biocatalysts
表征蛋白质动力学和催化功能之间的联系,以改进酶生物催化剂的设计
  • 批准号:
    RGPIN-2016-05557
  • 财政年份:
    2021
  • 资助金额:
    --
  • 项目类别:
    Discovery Grants Program - Individual
Catalytic Activation of Chemical Bonds between Quaternary Carbon Atoms
季碳原子间化学键的催化活化
  • 批准号:
    21K18987
  • 财政年份:
    2021
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Challenging Research (Exploratory)
Probing the correlation between tunneling and catalytic efficiency of H-transferring enzymes
探究氢转移酶的隧道效应和催化效率之间的相关性
  • 批准号:
    563274-2021
  • 财政年份:
    2021
  • 资助金额:
    --
  • 项目类别:
    University Undergraduate Student Research Awards
Characterizing the link between protein dynamics and catalytic function to improve the design of enzyme biocatalysts
表征蛋白质动力学和催化功能之间的联系,以改进酶生物催化剂的设计
  • 批准号:
    RGPIN-2016-05557
  • 财政年份:
    2020
  • 资助金额:
    --
  • 项目类别:
    Discovery Grants Program - Individual
Novel mechanism of integrase (IN) resistance to Dolutegravir through epistatic interactions between IN and the nucleocapsid and polypurine tract regions of HIV-1
通过 IN 与 HIV-1 核衣壳和多嘌呤束区域之间的上位相互作用,整合酶 (IN) 对 Dolutegravir 产生耐药性的新机制
  • 批准号:
    10348121
  • 财政年份:
    2019
  • 资助金额:
    --
  • 项目类别:
Novel mechanism of integrase (IN) resistance to Dolutegravir through epistatic interactions between IN and the nucleocapsid and polypurine tract regions of HIV-1
通过 IN 与 HIV-1 核衣壳和多嘌呤束区域之间的上位相互作用,整合酶 (IN) 对 Dolutegravir 产生耐药性的新机制
  • 批准号:
    10091399
  • 财政年份:
    2019
  • 资助金额:
    --
  • 项目类别:
Epigenetic deregulation and tumor progression due to the loss of a novel interaction between HDAC1 and BAP1 in uveal melanoma
由于葡萄膜黑色素瘤中 HDAC1 和 BAP1 之间新型相互作用的丧失导致表观遗传失调和肿瘤进展
  • 批准号:
    10238965
  • 财政年份:
    2019
  • 资助金额:
    --
  • 项目类别:
Novel mechanism of integrase (IN) resistance to Dolutegravir through epistatic interactions between IN and the nucleocapsid and polypurine tract regions of HIV-1
通过 IN 与 HIV-1 核衣壳和多嘌呤束区域之间的上位相互作用,整合酶 (IN) 对 Dolutegravir 产生耐药性的新机制
  • 批准号:
    10579840
  • 财政年份:
    2019
  • 资助金额:
    --
  • 项目类别:
Novel mechanism of integrase (IN) resistance to Dolutegravir through epistatic interactions between IN and the nucleocapsid and polypurine tract regions of HIV-1
通过 IN 与 HIV-1 核衣壳和多嘌呤束区域之间的上位相互作用,整合酶 (IN) 对 Dolutegravir 产生耐药性的新机制
  • 批准号:
    9803981
  • 财政年份:
    2019
  • 资助金额:
    --
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了