Allosteric impact of non-active-site mutations on enzymatic function
非活性位点突变对酶功能的变构影响
基本信息
- 批准号:10692526
- 负责人:
- 金额:$ 6.82万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2017
- 资助国家:美国
- 起止时间:2017-09-05 至 2024-06-30
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Antibiotic-resistant infections kill tens of thousands of Americans and cost our nation billions of
dollars every year. β-lactamase enzymes are one of the most common sources of resistance and
are capable of quickly evolving the ability to degrade new β-lactam antibiotics as they are
introduced. Surprisingly, many of the mutations that confer β-lactamases with new functions are
far from the enzyme's active site and have little effect on the structure of the active site, as
observed by x-ray crystallography. Such non-active site (NAS) mutations also appear frequently in
other contexts, such as the evolution of other forms of drug resistance and directed evolution
studies. Understanding how NAS mutations allosterically impact distant sites would provide a
basis for predicting new forms of drug resistance and designing allosteric drugs to combat
diseases like antibiotic-resistant infections. The objective of this proposal is to understand how
NAS mutations confer β- lactamases with activity against new substrates. A predictive
understanding of NAS mutations remains elusive because of the ruggedness of proteins' energy
landscapes and the great diversity of mechanisms that couple distant residues, including both
concerted structural changes and correlations between the dynamics of different residues. These
obstacles will be overcome by integrating novel computational methods with in vitro and in vivo
experiments to converge on a quantitative understanding of the full spectrum of correlated
fluctuations responsible for allosteric coupling. For example, the research team will apply new
methods they developed to facilitate comprehensive sampling of proteins' energy landscapes,
such as their FAST algorithm for leveraging Markov State Models (MSMs) to efficiently sample
conformations with pre-specified features. In Aim 1, these methods will be used to identify what
features of β-lactamase’s structure and dynamics give rise to new activities by comparing models
for variants with different activities against the antibiotic cefotaxime. In aim 2, new methods for
identifying both concerted structural changes and correlations between the dynamics of different
residues will be developed. These methods will be used to predict new sites where NAS mutations
can alter activities of β-lactamases. To test insights from each aim, mutations will be designed to
confer β- lactamases with new activities. Then experiments will be performed to test 1) whether
these mutations have the intended impact on the activities of β-lactamases and 2) whether the
designed variants are capable of protecting bacteria from the target antibiotic. Completion of this
work will result in a general framework for understanding allosteric communication that will serve
as a basis for future efforts to predict drug resistance, design new antibiotics that allosterically
inhibit their targets, and manipulate allostery in other systems.
抗生素耐药性感染杀死了成千上万的美国人,使我们的国家损失了数十亿美元。
美元每年。β-内酰胺酶是最常见的耐药性来源之一,
能够快速进化降解新β-内酰胺抗生素的能力,
介绍令人惊讶的是,许多赋予β-内酰胺酶新功能的突变是
远离酶的活性位点,对活性位点的结构几乎没有影响,
通过X射线晶体学观察。这种非活性位点(NAS)突变也经常出现在
其他背景下,如其他形式的耐药性和定向进化的演变
问题研究了解NAS突变如何变构影响远端位点将提供
预测新形式的耐药性和设计变构药物的基础,
比如抗药性感染本建议的目的是了解如何
NAS突变赋予β-内酰胺酶对新底物的活性。预测
由于蛋白质能量的不稳定性,对NAS突变的理解仍然是难以捉摸的
景观和耦合遥远残留物的机制的巨大多样性,包括两者
协调一致的结构变化和不同残留物动态之间的相关性。这些
通过将新的计算方法与体外和体内相结合,
实验收敛于相关的全光谱的定量理解
负责变构偶联的波动。例如,研究小组将应用新的
他们开发的方法,以促进全面采样蛋白质的能量景观,
例如他们的FAST算法,用于利用马尔可夫状态模型(MSM)来有效地采样
具有预定特征的构象。在目标1中,这些方法将用于确定
β-内酰胺酶的结构和动力学特征通过比较模型产生新的活性
用于对抗生素头孢噻肟具有不同活性的变体。在目标2中,
确定协调一致的结构变化和不同国家动态之间的相关性,
残留物将被开发。这些方法将用于预测NAS突变的新位点,
可以改变β-内酰胺酶的活性。为了测试每个目标的洞察力,突变将被设计成
赋予β-内酰胺酶新的活性。然后将进行实验以测试1)是否
这些突变对β-内酰胺酶的活性具有预期的影响,以及2)
设计的变体能够保护细菌免受目标抗生素的侵害。完成本
这项工作将产生一个理解变构通讯的总体框架,
作为未来努力预测耐药性的基础,设计新的抗生素,
抑制它们的目标,并操纵其他系统中的变构。
项目成果
期刊论文数量(17)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Mechanistic Basis for ATP-Dependent Inhibition of Glutamine Synthetase by Tabtoxinine-β-lactam.
- DOI:10.1021/acs.biochem.7b00838
- 发表时间:2018-01-09
- 期刊:
- 影响因子:2.9
- 作者:Patrick GJ;Fang L;Schaefer J;Singh S;Bowman GR;Wencewicz TA
- 通讯作者:Wencewicz TA
Choice of Adaptive Sampling Strategy Impacts State Discovery, Transition Probabilities, and the Apparent Mechanism of Conformational Changes.
- DOI:10.1021/acs.jctc.8b00500
- 发表时间:2018-11-13
- 期刊:
- 影响因子:5.5
- 作者:Zimmerman MI;Porter JR;Sun X;Silva RR;Bowman GR
- 通讯作者:Bowman GR
SARS-CoV-2 Nsp16 activation mechanism and a cryptic pocket with pan-coronavirus antiviral potential.
- DOI:10.1016/j.bpj.2021.03.024
- 发表时间:2021-07-20
- 期刊:
- 影响因子:3.4
- 作者:Vithani N;Ward MD;Zimmerman MI;Novak B;Borowsky JH;Singh S;Bowman GR
- 通讯作者:Bowman GR
Opening of a cryptic pocket in β-lactamase increases penicillinase activity.
- DOI:10.1073/pnas.2106473118
- 发表时间:2021-11-23
- 期刊:
- 影响因子:11.1
- 作者:Knoverek CR;Mallimadugula UL;Singh S;Rennella E;Frederick TE;Yuwen T;Raavicharla S;Kay LE;Bowman GR
- 通讯作者:Bowman GR
Enspara: Modeling molecular ensembles with scalable data structures and parallel computing.
Enspara:利用可扩展的数据结构和并行计算对分子整体进行建模。
- DOI:10.1063/1.5063794
- 发表时间:2019
- 期刊:
- 影响因子:0
- 作者:Porter,JR;Zimmerman,MI;Bowman,GR
- 通讯作者:Bowman,GR
{{
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 }}
Gregory R Bowman其他文献
Gregory R Bowman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('Gregory R Bowman', 18)}}的其他基金
Structural basis for ApoE4-induced Alzheimer's disease
ApoE4 诱导的阿尔茨海默病的结构基础
- 批准号:
10744482 - 财政年份:2021
- 资助金额:
$ 6.82万 - 项目类别:
MSMs, adaptive sampling, and data sharing on the cloud
MSM、自适应采样和云端数据共享
- 批准号:
10166370 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
ALLOSTERIC IMPACT OF NON-ACTIVE-SITE MUTATIONS ON ENZYMATIC FUNCTION
非活性位点突变对酶功能的变构影响
- 批准号:
10387558 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
ALLOSTERIC IMPACT OF NON-ACTIVE-SITE MUTATIONS ON ENZYMATIC FUNCTION
非活性位点突变对酶功能的变构影响
- 批准号:
9361418 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
ALLOSTERIC IMPACT OF NON-ACTIVE-SITE MUTATIONS ON ENZYMATIC FUNCTION
非活性位点突变对酶功能的变构影响
- 批准号:
9977221 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
ALLOSTERIC IMPACT OF NON-ACTIVE-SITE MUTATIONS ON ENZYMATIC FUNCTION
非活性位点突变对酶功能的变构影响
- 批准号:
10214633 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
ALLOSTERIC IMPACT OF NON-ACTIVE-SITE MUTATIONS ON ENZYMATIC FUNCTION
非活性位点突变对酶功能的变构影响
- 批准号:
9557495 - 财政年份:2017
- 资助金额:
$ 6.82万 - 项目类别:
相似国自然基金
The Heterogenous Impact of Monetary Policy on Firms' Risk and Fundamentals
- 批准号:
- 批准年份:2024
- 资助金额:万元
- 项目类别:外国学者研究基金项目
基于ImPACT方案的家长干预对孤独症谱系障碍儿童干预疗效及神经生物学机制研究
- 批准号:82301732
- 批准年份:2023
- 资助金额:30 万元
- 项目类别:青年科学基金项目
西方饮食通过“肠道菌群-Rspo1”轴促进肥胖与肠道吸收的机制研究
- 批准号:82370845
- 批准年份:2023
- 资助金额:48.00 万元
- 项目类别:面上项目
2型糖尿病胰岛β细胞功能调控新靶点IMPACT的功能及作用机制研究
- 批准号:81600598
- 批准年份:2016
- 资助金额:19.0 万元
- 项目类别:青年科学基金项目
基于IMPACT模型的社区慢性病干预效果的经济学评价研究
- 批准号:71303173
- 批准年份:2013
- 资助金额:21.0 万元
- 项目类别:青年科学基金项目
相似海外基金
Testing the genetic impact on the internal and external shape of teeth in non-human primates
测试遗传对非人类灵长类动物牙齿内部和外部形状的影响
- 批准号:
2341544 - 财政年份:2024
- 资助金额:
$ 6.82万 - 项目类别:
Standard Grant
Development of time-resolved rheological techniques using impact shear cells for study of phase transitions in non-Newtonian fluids
使用冲击剪切池开发时间分辨流变技术来研究非牛顿流体中的相变
- 批准号:
23K13243 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Impact of Alcohol Home Delivery on Access, Consumption, and Exposure to Marketing
酒类送货上门对获取、消费和营销曝光的影响
- 批准号:
10727955 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
Role of SIK3 in PKA/mTORC1 regulation of adipose browning
SIK3 在 PKA/mTORC1 调节脂肪褐变中的作用
- 批准号:
10736962 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
The space-time organization of sleep oscillations as potential biomarker for hypersomnolence
睡眠振荡的时空组织作为嗜睡的潜在生物标志物
- 批准号:
10731224 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
Identifying Metabolic and Psychosocial Antecedents and Characteristics of youth-onset Type 2 diabetes (IMPACT DM)
确定青年发病 2 型糖尿病 (IMPACT DM) 的代谢和心理社会因素和特征
- 批准号:
10584028 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
Determining the impact of ancestry on oropharyngeal cancer biology and treatment response.
确定血统对口咽癌生物学和治疗反应的影响。
- 批准号:
10562456 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
The non-invasive early detection of endometriosis
子宫内膜异位症的非侵入性早期检测
- 批准号:
10574971 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别:
Impact of Obesity on Chemotherapy-Induced Cytotoxicity: Immune Cells and Skeletal Muscle
肥胖对化疗引起的细胞毒性的影响:免疫细胞和骨骼肌
- 批准号:
10572695 - 财政年份:2023
- 资助金额:
$ 6.82万 - 项目类别: