Characterization of bacterial sensors using protein design
使用蛋白质设计表征细菌传感器
基本信息
- 批准号:9165148
- 负责人:
- 金额:$ 9万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2016
- 资助国家:美国
- 起止时间:2016-09-22 至 2018-08-31
- 项目状态:已结题
- 来源:
- 关键词:AddressAffectAmino AcidsAnti-Bacterial AgentsAntibiotic ResistanceAntibioticsApplications GrantsAreaAwardBacteriaBiological AssayBiologyBiophysicsCaliforniaCatalytic DomainCouplesCouplingCrystallizationCrystallographyDNADataDrug DesignDrug TargetingDrug resistanceEffectivenessEnvironmentEnzymesEquilibriumFaceFacultyFutureGene ExpressionGene ProteinsGoalsLaboratoriesLibrariesMapsMeasuresMembraneMentorsMethodsMicrofluidicsModificationMolecularMolecular ConformationPharmaceutical PreparationsPhasePhenotypePhosphoric Monoester HydrolasesPhosphotransferasesPopulationPredispositionPrincipal InvestigatorProkaryotic CellsProlinePropertyProtein EngineeringProteinsPublicationsReporterResearchResearch PersonnelResearch TrainingRoentgen RaysSan FranciscoScientistShapesSignal PathwaySignal TransductionStimulusStructureSystemTechnologyTemperatureTertiary Protein StructureTestingTrainingTransmembrane DomainUniversitiesVertebral columnVirulenceWorkX-Ray Crystallographyabstractingantimicrobialbacterial resistancebasecareercombinatorialconformational conversiondesigndimerexperienceflexibilityglobal healthnext generation sequencingprotein structure functionprotein-histidine kinaseresponsesensorskillssmall moleculesmall molecule inhibitor
项目摘要
Project Summary/Abstract
Two-component systems are major signaling pathways bacteria use to sense diverse stimuli such as
temperature, osmotic changes, and antibiotics and to initiate adaptive responses. In these systems the
histidine kinase (HK) detects the stimulus and relays the signal to its cognate response regulator, which alters
gene expression. As a postdoctoral scholar in Dr. William DeGrado's laboratory at the University of California,
San Francisco (UCSF), I have been designing protein constructs to study the structural and conformational
dynamics of HKs. Here, I propose to use an integrative experimental approach combining bacterial reporters,
enzymatic assays, x-ray structural studies, and droplet-based microfluidics technologies to understand how
conformational transitions in histidine kinases facilitate signal transduction. Recent HK structures suggest that
symmetry across the dimer interface is intricately related to catalytic state. The goal of this proposal is to
develop a molecular description of how structural signals induce symmetric to asymmetric conformational
transitions in the catalytic, cytoplasmic regions of HKs. I hypothesize that bistability of the dimer interface
`backbone' confers an essential conformational flexibility, which allows localized helical buckling to occur. The
consequence of this design is a transition from a continuous helical path along the backbone in the symmetric
state to a discontinuous path, which produces asymmetry. In Aim 1 I will examine how residue changes in the
buckling region of the backbone affects HK signaling. In Aim 2 I will use protein design to determine the
structural states associated with signaling in the cytoplasmic region of HKs. In Aim 3 I examine how signals
transmitted into the cytoplasmic region of HKs become modulated by coupling between effector domains and
the catalytic core. Completion of these aims will provide a molecular description of the structural and
conformational dynamics of HKs. A better understanding of the structural states and conformational changes
associated with signaling can inform structure-based design of small molecule inhibitors. By performing the
research in this proposal, I will increase my proficiency in protein design and biophysics while simultaneously
receiving strong training in X-ray crystallography, microfluidics, and high-throughput approaches to biology.
Expertise in these areas will better allow me to pursue my long-term scientific goals of using protein design as
a method to elucidate protein structure and function. The experience I will receive by working with my mentor
Dr. DeGrado and my collaborators during the K99 phase of the award will help me to become a stronger
scientist and better prepare me for a career as an independent researcher. The data and publications that
result from doing the work in this proposal will make me a stronger faculty candidate and help with future grant
applications. In general, completion of the research and training proposed in my application will provide me
with the skill set necessary for achieving my long-term career goal of becoming a principal investigator.
项目概要/摘要
双组分系统是细菌用来感知不同刺激的主要信号传导途径,例如
温度、渗透压变化和抗生素并启动适应性反应。在这些系统中
组氨酸激酶 (HK) 检测刺激并将信号传递给其同源反应调节器,从而改变
基因表达。作为加州大学William DeGrado博士实验室的博士后学者,
旧金山(UCSF),我一直在设计蛋白质构建体来研究结构和构象
香港的动态。在这里,我建议使用结合细菌报告基因的综合实验方法,
酶测定、X 射线结构研究和基于液滴的微流体技术,以了解如何
组氨酸激酶的构象转变促进信号转导。最近的香港结构表明
二聚体界面的对称性与催化状态密切相关。该提案的目标是
开发结构信号如何诱导对称到不对称构象的分子描述
HK 催化细胞质区域的转变。我假设二聚体界面的双稳定性
“主干”赋予了必要的构象灵活性,从而允许发生局部螺旋屈曲。这
这种设计的结果是从沿着对称结构主干的连续螺旋路径过渡
状态到不连续路径,从而产生不对称性。在目标 1 中,我将研究残留物如何变化
骨干的屈曲区域影响 HK 信号传输。在目标 2 中,我将使用蛋白质设计来确定
与 HK 细胞质区域信号传导相关的结构状态。在目标 3 中,我研究了如何发出信号
传输到 HK 细胞质区域的信号通过效应域和
催化核心。完成这些目标将提供结构和结构的分子描述。
HK 的构象动力学。更好地理解结构状态和构象变化
与信号传导相关的信息可以为小分子抑制剂的基于结构的设计提供信息。通过执行
在这个提案的研究中,我将提高我在蛋白质设计和生物物理学方面的熟练程度,同时
接受 X 射线晶体学、微流体学和高通量生物学方法方面的严格培训。
这些领域的专业知识将使我能够更好地追求我的长期科学目标,即利用蛋白质设计作为
一种阐明蛋白质结构和功能的方法。与我的导师一起工作我将获得的经验
DeGrado博士和我在K99阶段获奖的合作者将帮助我变得更强
科学家,让我更好地为独立研究员的职业生涯做好准备。数据和出版物
完成本提案中的工作的结果将使我成为一名更强大的教师候选人,并有助于获得未来的资助
应用程序。一般来说,完成我的申请中提出的研究和培训将为我提供
具备实现成为首席研究员的长期职业目标所需的技能。
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 }}
Nathan Schmidt其他文献
Nathan Schmidt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('Nathan Schmidt', 18)}}的其他基金
Role of the gut microbiota in shaping severity of malaria
肠道微生物群在影响疟疾严重程度中的作用
- 批准号:
9412115 - 财政年份:2017
- 资助金额:
$ 9万 - 项目类别:
Role of the gut microbiota in shaping severity of malaria
肠道微生物群在影响疟疾严重程度中的作用
- 批准号:
9307118 - 财政年份:2017
- 资助金额:
$ 9万 - 项目类别:
Role of the gut microbiota in shaping severity of malaria
肠道微生物群在影响疟疾严重程度中的作用
- 批准号:
10080707 - 财政年份:2017
- 资助金额:
$ 9万 - 项目类别:
相似海外基金
How Does Particle Material Properties Insoluble and Partially Soluble Affect Sensory Perception Of Fat based Products
不溶性和部分可溶的颗粒材料特性如何影响脂肪基产品的感官知觉
- 批准号:
BB/Z514391/1 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Training Grant
BRC-BIO: Establishing Astrangia poculata as a study system to understand how multi-partner symbiotic interactions affect pathogen response in cnidarians
BRC-BIO:建立 Astrangia poculata 作为研究系统,以了解多伙伴共生相互作用如何影响刺胞动物的病原体反应
- 批准号:
2312555 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Standard Grant
RII Track-4:NSF: From the Ground Up to the Air Above Coastal Dunes: How Groundwater and Evaporation Affect the Mechanism of Wind Erosion
RII Track-4:NSF:从地面到沿海沙丘上方的空气:地下水和蒸发如何影响风蚀机制
- 批准号:
2327346 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Standard Grant
Graduating in Austerity: Do Welfare Cuts Affect the Career Path of University Students?
紧缩毕业:福利削减会影响大学生的职业道路吗?
- 批准号:
ES/Z502595/1 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Fellowship
感性個人差指標 Affect-X の構築とビスポークAIサービスの基盤確立
建立个人敏感度指数 Affect-X 并为定制人工智能服务奠定基础
- 批准号:
23K24936 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Insecure lives and the policy disconnect: How multiple insecurities affect Levelling Up and what joined-up policy can do to help
不安全的生活和政策脱节:多种不安全因素如何影响升级以及联合政策可以提供哪些帮助
- 批准号:
ES/Z000149/1 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Research Grant
How does metal binding affect the function of proteins targeted by a devastating pathogen of cereal crops?
金属结合如何影响谷类作物毁灭性病原体靶向的蛋白质的功能?
- 批准号:
2901648 - 财政年份:2024
- 资助金额:
$ 9万 - 项目类别:
Studentship
Investigating how double-negative T cells affect anti-leukemic and GvHD-inducing activities of conventional T cells
研究双阴性 T 细胞如何影响传统 T 细胞的抗白血病和 GvHD 诱导活性
- 批准号:
488039 - 财政年份:2023
- 资助金额:
$ 9万 - 项目类别:
Operating Grants
New Tendencies of French Film Theory: Representation, Body, Affect
法国电影理论新动向:再现、身体、情感
- 批准号:
23K00129 - 财政年份:2023
- 资助金额:
$ 9万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
The Protruding Void: Mystical Affect in Samuel Beckett's Prose
突出的虚空:塞缪尔·贝克特散文中的神秘影响
- 批准号:
2883985 - 财政年份:2023
- 资助金额:
$ 9万 - 项目类别:
Studentship