Structural basis for cell surface siganling by a Gram-negative bacteria sigma-regulator
革兰氏阴性菌西格玛调节器细胞表面信号的结构基础
基本信息
- 批准号:10240569
- 负责人:
- 金额:$ 29万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2018
- 资助国家:美国
- 起止时间:2018-09-20 至 2024-08-31
- 项目状态:已结题
- 来源:
- 关键词:AdjuvantAffinityAntibiotic ResistanceAntibioticsBacteriaBacterial Antibiotic ResistanceBacterial InfectionsBacteriophagesBiological ModelsC-terminalCalorimetryCause of DeathCell surfaceCellsCellular AssayCenters for Disease Control and Prevention (U.S.)Cessation of lifeCircular Dichroism SpectroscopyComplexCyclic AMP Receptor ProteinDevelopmentDiarrheaDimerizationDiseaseDrug Delivery SystemsEnvironmentEquilibriumEquus caballusEscherichia coliFutureGastrointestinal tract structureGenetic TranscriptionGram-Negative BacteriaHealthHealth Care CostsHeavy MetalsHospitalsHumanIncidenceIronLeadLengthLength of StayLivestockMembraneMetalsMicrobial BiofilmsModernizationMolecular BiologyMolecular ConformationMulti-Drug ResistanceNatural ProductsNutrientPathogenicityPharmacologyPopulationPrevalencePrimary InfectionProcessPseudomonas aeruginosaPumpRecoveryRegulationReportingResearchResistanceResourcesRoentgen RaysRoleSiderophoresSigma FactorSignal TransductionSpecificityStimulusStomachStructureSystemTherapeuticTimeTitrationsToxic effectTranscriptional ActivationTranscriptional RegulationUp-RegulationVancomycinX-Ray Crystallographyantimicrobialbacteriocinbeta-Lactamsbiophysical techniquescarbapenem-resistant Enterobacteriaceaecombatcystic fibrosis infectioncystic fibrosis patientsdesigndisabilityefflux pumpexperimental studygastrointestinalhealth care settingsimprovedinterdisciplinary approachinterestmetal chelatormicroorganismmultidrug-resistant Pseudomonas aeruginosanext generationnovelnovel therapeuticspathogenpathogenic bacteriaperiplasmpressurepreventreceptorresistance mechanismsecondary infectionside effecttargeted deliveryuptake
项目摘要
PROJECT SUMMARY/ABSTRACT
The CDC recently released a report detailing antibiotic resistant threats in the US. Of particular
emphasis in the CDC report is the increased prevalence of multidrug-resistant, Gram-negative bacteria (MDR-
GNB) and the need to develop the next generation of antibiotics to combat them. All Gram-negative bacteria
rely on a set of homologous, yet highly-specific, outer membrane TonB-dependent transporters (TBDTs) to
import critical nutrients from their environment, especially metals like iron, which are bound by high-affinity,
metal chelating compounds called siderophores. Recent antibiotic developments have shown that
siderophore-antibiotic conjugates can be selectively targeted to specific bacteria, and that this delivery
mechanism overcomes several key antibiotic resistance mechanisms. A significant limitation of this delivery
system is the low expression levels of the TBDTs. However, a subset of these TBDTs controls their own
expression through a cell-surface signaling (CSS) process that up-regulates their own expression. The long-
term objective of this research is to understand the CSS regulatory process and manipulate TBDT expression
to enhance siderophore-antibiotic conjugate therapy for treatment of MDR-GNB infections. Research outlined
in this proposal will help elucidate the structural basis for CSS by a sigma-regulator. As a model system, the
pseudobactin BN7/8 transport system of Psuedomonas putida, which consists of the TBDT, PupB, the inner
membrane σ-regulator, PupR, and the cytoplasmic σ-factor, PupI, is being used. To accomplish this
proposal's objective the following three specific aims will be pursued: 1) establish that PupR anti-σ-factor
domain dimerization influences transcriptional activation by PupI, 2) identify the structural determinants and
delineate the role of the PupR:PupB periplasmic interactions on the stability of the PupR periplasmic C-
terminal CSS domain (CCSSD), and 3) determine changes in the full-length PupB:PupR CCSSD complex in
the presence and absence of its cognate siderophore, pseudobactin BN7/8. These aims will be accomplished
using a multidisciplinary approach; including X-ray crystallography, small-angle X-ray scattering, molecular
biology, cellular assays, and biophysical techniques such as isothermal titration calorimetry and circular
dichroism spectroscopy. This research will provide critical structural information about a σ-regulator; explain
how it interacts with a σ-factor at the inner membrane, and the extent to which periplasmic conformational
changes between the TBDT and σ-regulator lead to proteolytic degradation that is important for controlling
transcriptional activation.
项目概要/摘要
疾病预防控制中心最近发布了一份报告,详细介绍了美国的抗生素耐药性威胁。特别是
CDC 报告强调的是多重耐药革兰氏阴性菌(MDR-
GNB)以及开发下一代抗生素来对抗它们的需要。所有革兰氏阴性细菌
依赖于一组同源但高度特异性的外膜 TonB 依赖性转运蛋白 (TBDT)
从环境中输入关键的营养物质,特别是像铁这样的金属,它们通过高亲和力结合在一起,
称为铁载体的金属螯合化合物。最近抗生素的发展表明
铁载体-抗生素缀合物可以选择性地靶向特定细菌,并且这种递送
机制克服了几个关键的抗生素耐药机制。这种交付的一个重大限制
系统中TBDTs的表达水平较低。然而,这些 TBDT 的一个子集控制着它们自己的
通过细胞表面信号传导(CSS)过程上调其自身表达。长-
本研究的长期目标是了解 CSS 调控过程并操纵 TBDT 表达
增强铁载体-抗生素结合疗法治疗 MDR-GNB 感染。研究概述
该提案中的内容将有助于通过 sigma 调节器阐明 CSS 的结构基础。作为一个模型系统,
恶臭假单胞菌的假杆菌素BN7/8转运系统,由TBDT、PupB、内
使用膜 σ 调节因子 PupR 和细胞质 σ 因子 PupI。为了实现这一点
提案的目标 将追求以下三个具体目标: 1) 确定 PupR 抗 σ 因子
结构域二聚化影响 PupI 的转录激活,2) 识别结构决定因素和
描述 PupR:PupB 周质相互作用对 PupR 周质 C-稳定性的作用
终端 CSS 域 (CCSSD),以及 3) 确定全长 PupB:PupR CCSSD 复合体的变化
其同源铁载体假杆菌素 BN7/8 的存在和不存在。这些目标将会实现
采用多学科方法;包括X射线晶体学、小角X射线散射、分子
生物学、细胞测定和生物物理技术,例如等温滴定量热法和循环法
二色性光谱。这项研究将提供有关 σ 调节器的关键结构信息;解释
它如何与内膜的 σ 因子相互作用,以及周质构象的程度
TBDT 和 σ 调节剂之间的变化会导致蛋白水解降解,这对于控制蛋白水解很重要
转录激活。
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 }}
Christopher L Colbert其他文献
Christopher L Colbert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('Christopher L Colbert', 18)}}的其他基金
Analytical ultracentrifuge with absorbance and interference optics.
具有吸光度和干涉光学器件的分析超速离心机。
- 批准号:
10177341 - 财政年份:2021
- 资助金额:
$ 29万 - 项目类别:
Structural basis for cell surface siganling by a Gram-negative bacteria sigma-regulator
革兰氏阴性菌西格玛调节器细胞表面信号的结构基础
- 批准号:
9789675 - 财政年份:2018
- 资助金额:
$ 29万 - 项目类别:
Structural basis for cell surface siganling by a Gram-negative bacteria sigma-regulator
革兰氏阴性菌西格玛调节器细胞表面信号的结构基础
- 批准号:
10004679 - 财政年份:2018
- 资助金额:
$ 29万 - 项目类别:
Structural basis for cell surface siganling by a Gram-negative bacteria sigma-regulator
革兰氏阴性菌西格玛调节器细胞表面信号的结构基础
- 批准号:
10387865 - 财政年份:2018
- 资助金额:
$ 29万 - 项目类别:
Mechanism of inner membrane sigma-regulator function in Gram-negative bacteria
革兰氏阴性菌内膜西格玛调节功能的机制
- 批准号:
9316212 - 财政年份:2015
- 资助金额:
$ 29万 - 项目类别:
相似海外基金
Construction of affinity sensors using high-speed oscillation of nanomaterials
利用纳米材料高速振荡构建亲和传感器
- 批准号:
23H01982 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Affinity evaluation for development of polymer nanocomposites with high thermal conductivity and interfacial molecular design
高导热率聚合物纳米复合材料开发和界面分子设计的亲和力评估
- 批准号:
23KJ0116 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Grant-in-Aid for JSPS Fellows
Platform for the High Throughput Generation and Validation of Affinity Reagents
用于高通量生成和亲和试剂验证的平台
- 批准号:
10598276 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Development of High-Affinity and Selective Ligands as a Pharmacological Tool for the Dopamine D4 Receptor (D4R) Subtype Variants
开发高亲和力和选择性配体作为多巴胺 D4 受体 (D4R) 亚型变体的药理学工具
- 批准号:
10682794 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Collaborative Research: DESIGN: Co-creation of affinity groups to facilitate diverse & inclusive ornithological societies
合作研究:设计:共同创建亲和团体以促进多元化
- 批准号:
2233343 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Standard Grant
Collaborative Research: DESIGN: Co-creation of affinity groups to facilitate diverse & inclusive ornithological societies
合作研究:设计:共同创建亲和团体以促进多元化
- 批准号:
2233342 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Standard Grant
Molecular mechanisms underlying high-affinity and isotype switched antibody responses
高亲和力和同种型转换抗体反应的分子机制
- 批准号:
479363 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Operating Grants
Deconstructed T cell antigen recognition: Separation of affinity from bond lifetime
解构 T 细胞抗原识别:亲和力与键寿命的分离
- 批准号:
10681989 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
CAREER: Engineered Affinity-Based Biomaterials for Harnessing the Stem Cell Secretome
职业:基于亲和力的工程生物材料用于利用干细胞分泌组
- 批准号:
2237240 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Continuing Grant
ADVANCE Partnership: Leveraging Intersectionality and Engineering Affinity groups in Industrial Engineering and Operations Research (LINEAGE)
ADVANCE 合作伙伴关系:利用工业工程和运筹学 (LINEAGE) 领域的交叉性和工程亲和力团体
- 批准号:
2305592 - 财政年份:2023
- 资助金额:
$ 29万 - 项目类别:
Continuing Grant