Chemogenetic control of kinase and phosphatase activity by modulating autoinhibition
通过调节自抑制对激酶和磷酸酶活性进行化学遗传学控制
基本信息
- 批准号:10195182
- 负责人:
- 金额:$ 23.37万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-04-01 至 2023-03-31
- 项目状态:已结题
- 来源:
- 关键词:Active SitesAffinityAnimalsBehaviorBindingBiomedical ResearchCREB1 geneCa(2+)-Calmodulin Dependent Protein KinaseCalcineurinCalciumCalmodulinCatalytic DomainCell Surface ReceptorsCell TherapyCellsCellular biologyChemicalsClinicalComplementComplexDissociationDrug ControlsDrug usageElementsEngineeringEnzyme ActivationEnzymesGenetic TranscriptionHeterodimerizationHoloenzymesIL2 geneInterleukin-2KineticsLengthLymphocyteMediatingMethodsOutcomePathway interactionsPeptidesPerformancePermeabilityPharmaceutical PreparationsPharmacologyPhosphoric Monoester HydrolasesPhosphotransferasesPositioning AttributeProtein phosphataseProteinsRegulationRoleSignal PathwaySignal TransductionSignaling ProteinStructureSystemT-LymphocyteTechniquesTestingWorkbasecalcineurin phosphatasedesigndimerengineered T cellsenzyme activityexperimental studygene therapyimprovedin vivoinhibitor/antagonistinnovationinterestnext generationnovelpreventprotein functionsmall moleculetool
项目摘要
ABSTRACT
Signaling enzymes such as kinases and phosphatases control multiple aspects of cellular differentiation and
behavior, and are especially important in transducing signals from cell surface receptors to changes in cell fate
or function. The ability to activate signaling proteins of interest using validated cell-permeable drugs would be
immensely useful for studying the functions of these proteins in cells or animals, and could provide much-needed
control over gene and cell therapies.
Here, we propose a novel method for conferring chemical control over kinases and phosphatases based on
drug-induced displacement of a tethered autoinhibitory domain (AID) from the active site. We will test and
validate this method using the phosphatase calcineurin (CaN) and calcium/calmodulin kinase IV (CaMKIV), two
enzymes that are natively inhibited by an AID and activated by a mechanism involving AID dissociation. In our
method, we will use fused heterodimerizing elements to position the AIDs near the enzyme active site, then use
small-molecule drugs to disrupt this interaction and displace the AIDs from the active site. We will carry out the
following specific aims: (1) Creating drug-activated CaN using a chemically-dissociable autoinhibitory peptide,
(2) Creating drug-activated CaMKIV using a chemically-dissociable autoinhibitory peptide, and (3) Examining
roles of CaN and CaMKIV in IL-2 transcription in T cells using drug-activated proteins.
Our design has several unique and innovative features. The single-chain design should improve reliability and
reduce complexity over multi-component systems. The ability to rationally modulate linker length, heterodimer
affinity, and AID-enzyme affinity provides multiple avenues for construct optimization. Multiple chemically
dissociable interactions are known, allowing for multiplexed drug-controllable proteins. Finally, given that
intramolecular AIDs should be low rather than high affinity, peptide inhibitors can be selected or designed for
signaling enzymes that lack native AIDs. Our method of protein control by drug-induced displacement of an
autoinhibitory domain should thus be uniquely useful, robust, and generalizable.
抽象的
信号酶如激酶和磷酸酶控制细胞分化的多个方面
行为,并且对于将细胞表面受体的信号转导至细胞命运的变化尤其重要
或函数。使用经过验证的细胞渗透性药物激活感兴趣的信号蛋白的能力将是
对于研究细胞或动物中这些蛋白质的功能非常有用,并且可以提供急需的
控制基因和细胞疗法。
在这里,我们提出了一种基于化学控制激酶和磷酸酶的新方法
药物诱导的束缚自抑制结构域 (AID) 从活性位点移位。我们将测试并
使用磷酸酶钙调神经磷酸酶 (CaN) 和钙/钙调蛋白激酶 IV (CaMKIV) 验证此方法,两个
天然被 AID 抑制并通过涉及 AID 解离的机制激活的酶。在我们的
方法,我们将使用融合的异二聚化元件将 AID 定位在酶活性位点附近,然后使用
小分子药物可以破坏这种相互作用并从活性位点取代 AID。我们将开展
以下具体目标:(1) 使用化学可解离的自抑制肽创建药物激活的 CaN,
(2) 使用化学解离的自抑制肽创建药物激活的 CaMKIV,以及 (3) 检查
使用药物激活蛋白研究 CaN 和 CaMKIV 在 T 细胞 IL-2 转录中的作用。
我们的设计有几个独特和创新的特点。单链设计应提高可靠性和
降低多组件系统的复杂性。合理调节接头长度、异二聚体的能力
亲和力和 AID-酶亲和力为构建体优化提供了多种途径。多重化学
可解离的相互作用是已知的,允许多重药物可控蛋白质。最后,考虑到
分子内AID应该是低亲和力而不是高亲和力,可以选择或设计肽抑制剂
缺乏天然 AID 的信号酶。我们通过药物诱导的位移来控制蛋白质的方法
因此,自抑制域应该是独特有用的、稳健的和可推广的。
项目成果
期刊论文数量(0)
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科研奖励数量(0)
会议论文数量(0)
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Michael Z. Lin其他文献
Synaptic basis of feature selectivity in hippocampal neurons
海马神经元特征选择性的突触基础
- DOI:
10.1038/s41586-024-08325-9 - 发表时间:
2024-12-18 - 期刊:
- 影响因子:48.500
- 作者:
Kevin C. Gonzalez;Adrian Negrean;Zhenrui Liao;Satoshi Terada;Guofeng Zhang;Sungmoo Lee;Katalin Ócsai;Balázs J. Rózsa;Michael Z. Lin;Franck Polleux;Attila Losonczy - 通讯作者:
Attila Losonczy
An optimized luciferin formulation for NanoLuc-based in vivo bioluminescence imaging
用于基于 NanoLuc 的体内生物发光成像的优化荧光素制剂
- DOI:
10.1038/s41598-025-97366-9 - 发表时间:
2025-04-15 - 期刊:
- 影响因子:3.900
- 作者:
Chao Gao;Yan Wu;Connor Fitzgerald;Hui Wang;Tim Ugo;Tetsuo Uyeda;Wenhui Zhou;Yichi Su;Thomas A. Kirkland;Michael Z. Lin - 通讯作者:
Michael Z. Lin
Functional and Structural Characterization of A New Monomeric Far-Red Fluorescent Protein
- DOI:
10.1016/j.bpj.2009.12.1158 - 发表时间:
2010-01-01 - 期刊:
- 影响因子:
- 作者:
Michael Z. Lin;Michael R. McKeown;Ho Leung Ng;Tom Alber;Roger Y. Tsien - 通讯作者:
Roger Y. Tsien
On the cutting edge: protease-based methods for sensing and controlling cell biology
处于前沿:基于蛋白酶的细胞生物学传感与控制方法
- DOI:
10.1038/s41592-020-0891-z - 发表时间:
2020-07-13 - 期刊:
- 影响因子:32.100
- 作者:
H. Kay Chung;Michael Z. Lin - 通讯作者:
Michael Z. Lin
Pharmacodynamics of Akt drugs revealed by a kinase-modulated bioluminescent indicator
一种激酶调节的生物发光指示剂揭示的 Akt 药物的药效学
- DOI:
10.1038/s41589-025-01846-y - 发表时间:
2025-02-11 - 期刊:
- 影响因子:13.700
- 作者:
Yan Wu;Chenzhou Hao;Chao Gao;Matt Hageman;Sungmoo Lee;Thomas A. Kirkland;Nathanael S. Gray;Yichi Su;Michael Z. Lin - 通讯作者:
Michael Z. Lin
Michael Z. Lin的其他文献
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{{ truncateString('Michael Z. Lin', 18)}}的其他基金
Development of selective and potent protease inhibitors for corona and other pandemic viruses
开发针对冠状病毒和其他大流行病毒的选择性有效蛋白酶抑制剂
- 批准号:
10514273 - 财政年份:2022
- 资助金额:
$ 23.37万 - 项目类别:
The power of positivity: a novel class of voltage indicators for high-fidelity brain activity imaging
积极性的力量:用于高保真大脑活动成像的新型电压指示器
- 批准号:
10294164 - 财政年份:2021
- 资助金额:
$ 23.37万 - 项目类别:
Bioluminescent indicators for noninvasive imaging of acetylcholine release
用于乙酰胆碱释放无创成像的生物发光指示器
- 批准号:
10196839 - 财政年份:2021
- 资助金额:
$ 23.37万 - 项目类别:
Chemogenetic control of kinase and phosphatase activity by modulating autoinhibition
通过调节自抑制对激酶和磷酸酶活性进行化学遗传学控制
- 批准号:
10371123 - 财政年份:2021
- 资助金额:
$ 23.37万 - 项目类别:
Noninvasive bioluminescent imaging of neuronal activity in freely behaving animals
自由行为动物神经元活动的无创生物发光成像
- 批准号:
9906190 - 财政年份:2019
- 资助金额:
$ 23.37万 - 项目类别:
Protein voltage sensors: kilohertz imaging of neural dynamics in behaving animals
蛋白质电压传感器:行为动物神经动力学的千赫兹成像
- 批准号:
8827201 - 财政年份:2014
- 资助金额:
$ 23.37万 - 项目类别:
Optogenetics for all: A general method for optical control of protein activity
所有人的光遗传学:蛋白质活性光学控制的通用方法
- 批准号:
8896827 - 财政年份:2013
- 资助金额:
$ 23.37万 - 项目类别:
Optogenetics for all: A general method for optical control of protein activity
所有人的光遗传学:蛋白质活性光学控制的通用方法
- 批准号:
9132820 - 财政年份:2013
- 资助金额:
$ 23.37万 - 项目类别:
Optogenetics for all: A general method for optical control of protein activity
所有人的光遗传学:蛋白质活性光学控制的通用方法
- 批准号:
8564060 - 财政年份:2013
- 资助金额:
$ 23.37万 - 项目类别:
A Molecular Tag for Drug-Regulated Synthesis of Specific Proteins
用于药物调控合成特定蛋白质的分子标签
- 批准号:
8733708 - 财政年份:2011
- 资助金额:
$ 23.37万 - 项目类别:
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