Structural consequences of PKC-dependent phosphorylation of Kv7.2
Kv7.2 PKC 依赖性磷酸化的结构后果
基本信息
- 批准号:10609077
- 负责人:
- 金额:$ 12.5万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-05-01 至 2024-08-31
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalAffectAffinityAwardBindingBinding SitesBiological AssayCa(2+)-Calmodulin Dependent Protein KinaseCalmodulinCareer MobilityCell Culture TechniquesCryoelectron MicroscopyDataDiseaseElectrophysiology (science)EncephalopathiesEnvironmentEpilepsyEventFinancial SupportFoundationsFunctional disorderFutureG-Protein-Coupled ReceptorsGTP-Binding ProteinsHealthHumanInterdisciplinary StudyIon ChannelIsotope LabelingMapsMolecularMuscarinic M1 ReceptorMutationNeonatalPainPharmacologyPhasePhosphatidylinositol 4,5-DiphosphatePhosphatidylinositolsPhosphorylationPhysiologyPlayProtein IsoformsProtein KinaseProtein Kinase CRegulationReportingResearchResourcesRoleSeizuresSerineSignal TransductionSignaling MoleculeSiteStrokeStructureTechniquesTestingTrainingTraumatic Brain InjuryX-Ray CrystallographyXenopusalpha helixbiophysical techniquescareercofactorimprovedinnovationmutantneuronal excitabilitynoveloptogeneticspreventprogramsstoichiometrystructural biologysupportive environment
项目摘要
Project Summary
M channels are critical for regulating the excitability of neurons. Dysfunction of M channel activity can
cause epilepsy. While M channels have been intensely studied, the interplay of several G-protein-regulated
signaling cofactors on these channels is still poorly understood. M channels are hetero-tetrameric pore
structures formed by the combination of subunits Kv7.2-5. Over 80 mutations have been mapped to the Kv7.2
subunit, being a primary cause of neonatal epilepsy. Many of these mutations lie within the binding domains for
at least three critical signaling cofactors: calmodulin (CaM), phosphatidylinositol 4,5-bisphosphate (PIP2) and
protein kinase (PKC). How PIP2 and CaM binding to Kv7.2 harmonize to fine tune channel activity is obscure.
Moreover, the role played by PKC in tuning this binding is obscure. My preliminary data shows that PIP2 and
CaM may simultaneously bind the B helix with phosphorylation tempering this binding. My NMR studies so far
show that phosphorylation reconfigures the apoCaM-B helix interaction, suggesting the interplay between
these cofactors has significant impact on channel structure. I will test the overarching hypothesis that
phosphorylation at S520 and S527 fine-tunes the ability of PIP2 and CaM to bind to Kv7.2 and control M
channel activity. This hypothesis will be tested using three aims and will provide mechanistic understanding
of how phosphorylation, and dependent PIP2 and CaM binding affects the structure of Kv7.2 to control channel
activity. In Aim 1, I will use advanced 3D and 4D NMR to resolve the solution structure of purified Kv7.2 C
terminus. Aim 2 will use these NMR spectra to define the affinity of PIP2 to Kv7.2 and describe the
stoichiometry and mode of binding between PIP2 and the multiple sites on Kv7.2. Aim 3 will elaborate how
phosphorylation within the B helix directs the interplay between CaM and PIP2 binding to Kv7.2. The proposed
study is innovative because it will address a longstanding question about how and where PIP2 binds Kv7.2,
and will elaborate how phosphorylation directs the interplay between CaM and PIP2 as they bind Kv7.2. My
training in advanced 3D and 4D NMR will be critical for my career advancement as I plan to use this rigorous
technique throughout my career. The rich resources and supportive environment at UT Health combined with
my expertise in biophysical methods on ion channels makes me the ideal candidate to study the mechanisms
underlying the regulation of M channel activity. The MOSAIC career award will provide valuable financial
support to help me begin my multidisciplinary research program focused on elucidating the molecular
mechanisms of ion channel regulation.
项目摘要
M通道是调节神经元兴奋性的关键。M通道活动功能障碍可
导致癫痫。虽然M通道已经得到了深入的研究,但几种G蛋白调节的相互作用
这些通道上的信号辅助因子仍然知之甚少。M孔道为异四聚体孔道
Kv7.2-5亚基组合形成的结构。已有80多个突变被映射到Kv7.2
亚单位,是新生儿癫痫的主要原因。这些突变中的许多都存在于
至少三个关键的信号辅助因子:钙调蛋白(CaM)、磷脂酰肌醇4,5-二磷酸(PIP2)和
蛋白激酶(PKC)。PIP2和CaM与Kv7.2的结合如何协调以微调通道活动尚不清楚。
此外,PKC在调节这种结合方面所起的作用还不清楚。我的初步数据显示,PIP2和
CaM可以同时与B螺旋结合,并通过磷酸化来缓和这种结合。到目前为止我的核磁共振研究
表明磷酸化重新配置apoCaM-B螺旋相互作用,提示
这些共同因素对渠道结构有显著影响。我将测试最重要的假设
S520和S527的磷酸化微调PIP2和CaM结合Kv7.2和控制M的能力
通道活动。这一假设将通过三个目标进行检验,并将提供机械性的理解
以及依赖PIP2和CaM结合如何影响Kv7.2的结构以控制通道
活动。在目标1中,我将使用先进的3D和4D核磁共振来解析纯化的Kv7.2 C的溶液结构
终点站。AIM 2将使用这些核磁共振谱来定义PIP2对Kv7.2的亲和力,并描述
化学计量学和PIP2与Kv7.2上多个位点的结合方式。目标3将详细说明如何
B螺旋内的磷酸化引导CaM和PIP2与Kv7.2结合的相互作用。建议数
这项研究具有创新性,因为它将解决一个长期存在的问题,即PIP2如何以及在哪里绑定Kv7.2,
并将详细阐述磷酸化如何指导CaM和PIP2之间的相互作用,因为它们结合Kv7.2。我的
高级3D和4D核磁共振方面的培训对我的职业发展至关重要,因为我计划使用这一严格的
技术贯穿了我的职业生涯。UT Health丰富的资源和支持环境与
我在离子通道的生物物理方法方面的专业知识使我成为研究这种机制的理想人选。
这是M通道活动调节的基础。马赛克职业奖将提供宝贵的资金
支持我开始我的多学科研究计划,重点是阐明分子
离子通道调节机制。
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 }}
Crystal Rae Archer其他文献
Crystal Rae Archer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('Crystal Rae Archer', 18)}}的其他基金
Structural consequences of PKC-dependent phosphorylation of Kv7.2
Kv7.2 PKC 依赖性磷酸化的结构后果
- 批准号:
10429142 - 财政年份:2022
- 资助金额:
$ 12.5万 - 项目类别:
Biophysical interactions of PIP2 and calmodulin with KCNQ (Kv7) K+ ion channels
PIP2 和钙调蛋白与 KCNQ (Kv7) K 离子通道的生物物理相互作用
- 批准号:
8838438 - 财政年份:2015
- 资助金额:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 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:
$ 12.5万 - 项目类别:
Studentship