Defining protein:protein interactions for the regulation of renal V-ATPase function: role in expression, assembly and trafficking.
定义蛋白质:调节肾 V-ATP 酶功能的蛋白质相互作用:在表达、组装和运输中的作用。
基本信息
- 批准号:10207619
- 负责人:
- 金额:$ 54.17万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-09-01 至 2024-06-30
- 项目状态:已结题
- 来源:
- 关键词:ATP phosphohydrolaseAcid-Base EquilibriumAcidosisAcidsAddressAffectAmino AcidsBindingBiologicalBloodCell Culture TechniquesCell membraneCell surfaceCellsComplexCuesCyclic AMPDataDegradation PathwayDiseaseDistalDown-RegulationDrug DesignEpithelial CellsEventExocytosisFamilyFundingFutureGene ExpressionGenetic TranscriptionGoalsHoloenzymesHomologous GeneIn VitroIncentivesIntercalated CellIntercalated DuctIntracellular TransportIntuitionKidneyKnock-outKnockout MiceKnowledgeLifeLinkMalignant NeoplasmsMapsMissionMolecularMusMutagenesisMutationOrganPeptidesPharmaceutical PreparationsPlayProcessPropertyProteinsProton PumpProtonsPublic HealthPumpRecyclingRegulationRegulatory PathwayRenal tubular acidosisResearchRoleSiteTechniquesTranslationsUnited States National Institutes of HealthVesicleVirus DiseasesWorkYeastsbasebonecell typedesigndisease-causing mutationexperimental studyextracellulargenetic regulatory proteinhuman diseaseinnovationinsightkidney cellknock-downmouse modelnewsnovelprotein protein interactionrenal epitheliumresponsespatiotemporaltraffickingtranscriptome sequencingvacuolar H+-ATPasevesicle transport
项目摘要
PROJECT SUMMARY
Despite its central role in extracellular acidification in the kidney and other organs, as well as in critical
intracellular processes, the regulation of proton-pumping ATPase (V-ATPase) activity at the molecular level is poorly
understood. During the prior funding period, two proteins that associate strongly with the V-ATPase to regulate its
function were identified – Ncoa7 and Dmxl1. The overall objective of this proposal is to determine the mechanisms
by which they interact with the V-ATPase to regulate proton secretion by the kidney, thereby maintaining systemic
acid/base balance. The long-term goal is to develop strategies (including drug and peptide design) for the regulation
of acidification processes that are inappropriately up- or downregulated not only in the kidney, but also in diseases
affecting other cells and organs. Ncoa7 null mice have markedly decreased V-ATPase subunit expression in collecting
duct intercalated cells (ICs) resulting in distal RTA, while Dmxl1 knockdown in renal epithelial cells in vitro causes
deficient intracellular vesicle acidification to the same degree as knockdown of bone-fide V-ATPase subunits. Aim 1
will determine the mechanism by which Ncoa7 regulates V-ATPase subunit expression, focusing on translational,
transcriptional and degradation pathways in WT and Ncoa7 knockout mice. The V-ATPase-binding sequence of Ncoa7
will be identified by protein interaction and mutagenesis studies, and its role in V-ATPase-dependent acidification
events will be determined. Aim 2 will address the novel hypothesis that V-ATPase exocytosis and accumulation on
the plasma membrane of ICs in response to systemic acid/base cues requires, counter-intuitively, a partial disassembly
of the large, sterically hindering V-ATPase holoenzymes that coat intracellular transport vesicles. The working
hypothesis is that Dmxl1, a homolog of the Rav1p yeast V-ATPase assembly protein, coordinates V-ATPase
assembly/disassembly with V-ATPase recycling to and from the plasma membrane, which together regulate V-ATPase
activity and proton secretion in kidney ICs and other cells. The functionally important V-ATPase-binding sequence of
Dmxl1 will also be identified by protein interaction and mutagenesis studies. Thus, a major innovative aspect of our
proposed studies is the concept that two newly-identified V-ATPase interacting proteins are involved in the
regulation of V-ATPase function at the “upstream” expression level and the “downstream” assembly level. Both
Aims 1 and 2 make use of integrated cell and molecular techniques in conjunction with genetically modified mouse
models, isolated ICs and renal cell cultures in vitro, and interaction domain studies using purified proteins and specific
subdomains. The proposed research is significant: a) because it will allow the field to move forward not at the most
basic cellular level by elucidating new V-ATPase dependent acidification regulatory pathways, and b) because the
interaction site analysis will inform the future design of drugs and/or cell permeant biologics to up- or down-regulate
V-ATPase activity in states of inappropriate hyperactivation (e. g., in many cancers, viral infection) or downregulation
(such as dRTA).
项目总结
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)
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Dennis Brown其他文献
Dennis Brown的其他文献
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{{ truncateString('Dennis Brown', 18)}}的其他基金
Cell Biology of Vasopressin-induced Water Channels-Research Supplement
加压素诱导的水通道的细胞生物学-研究补充
- 批准号:
10835229 - 财政年份:2023
- 资助金额:
$ 54.17万 - 项目类别:
An Open-Labeled, Single Arm Phase 2 Efficacy and Safety Study of REM-001 Photodynamic Therapy (PDT) for Treatment of Cutaneous Metastatic Breast Cancer (CMBC)
REM-001 光动力疗法 (PDT) 治疗皮肤转移性乳腺癌 (CMBC) 的开放标记单臂 2 期疗效和安全性研究
- 批准号:
10699535 - 财政年份:2023
- 资助金额:
$ 54.17万 - 项目类别:
HD Upgrade to a Nikon A1R Confocal Imaging Platform
高清升级至尼康 A1R 共焦成像平台
- 批准号:
10415591 - 财政年份:2022
- 资助金额:
$ 54.17万 - 项目类别:
Defining protein:protein interactions for the regulation of renal V-ATPase function: role in expression, assembly and trafficking.
定义蛋白质:调节肾 V-ATP 酶功能的蛋白质相互作用:在表达、组装和运输中的作用。
- 批准号:
10670311 - 财政年份:2019
- 资助金额:
$ 54.17万 - 项目类别:
Defining protein:protein interactions for the regulation of renal V-ATPase function: role in expression, assembly and trafficking.
定义蛋白质:调节肾 V-ATP 酶功能的蛋白质相互作用:在表达、组装和运输中的作用。
- 批准号:
10454931 - 财政年份:2019
- 资助金额:
$ 54.17万 - 项目类别:
A Zeiss LSM800 confocal microscope with Airyscan
配备 Airyscan 的 Zeiss LSM800 共焦显微镜
- 批准号:
9075249 - 财政年份:2016
- 资助金额:
$ 54.17万 - 项目类别:
Cell biology of vasopressin-induced water channels
加压素诱导的水通道的细胞生物学
- 批准号:
10005038 - 财政年份:2012
- 资助金额:
$ 54.17万 - 项目类别:
Cell Biology of Vasopressin-induced Water Channels
加压素诱导的水通道的细胞生物学
- 批准号:
10652774 - 财政年份:2012
- 资助金额:
$ 54.17万 - 项目类别:
Cell biology of vasopressin-induced water channels
加压素诱导的水通道的细胞生物学
- 批准号:
9176186 - 财政年份:2012
- 资助金额:
$ 54.17万 - 项目类别:
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