Membrane-Mediated Interactions of the Bcl-xL/Bid/Bax Triad of Apoptotic Regulators
Membrane-Mediated Interactions of the Bcl-xL/Bid/Bax Triad of Apoptotic Regulators
批准号:
10335236
负责人:
ALEXEY LADOKHIN
金额:
$37.43万
依托单位国家:
美国
项目类别:
财政年份:
2019
资助国家:
美国
项目状态:
已结题
起止时间:
2019-02-01 至 2024-01-31
关键词:
ApoptosisApoptoticAutoimmunityBCL-2 ProteinBCL2 geneBCL2L1 geneCell DeathCell SurvivalCell physiologyComplementComputer SimulationDataDegenerative DisorderDevelopmentDiphtheria ToxinElectron Spin Resonance SpectroscopyElectrostaticsEnvironmentFluorescenceFluorescence Resonance Energy TransferFutureHyperactivityImmunologic Deficiency SyndromesKnowledgeLabelLipid BilayersLipidsMalignant NeoplasmsMapsMediatingMembraneModelingMolecularMolecular ConformationMolecular Mechanisms of ActionNerve DegenerationOuter Mitochondrial MembranePathway interactionsPhysiologicalPopulationProcessProtein ConformationProtein FamilyProteinsRegulationSiteSystemTherapeuticThermodynamicsTissuesTriad Acrylic Resincancer cellexperienceinsightmechanical propertiesmolecular dynamicsrecruittherapeutic development
中文摘要
总结
细胞凋亡对组织中细胞群的正常发育和功能至关重要,其异常增殖和凋亡是细胞凋亡的重要组成部分。
调节对于变性疾病和癌症具有重要意义。触发的关键步骤
细胞凋亡是线粒体外膜(MOMP)的透化。这个过程是紧密的
由Bcl-2蛋白家族调节,其被细分为促凋亡(例如,Bax),抗凋亡
(e.g., Bcl-xL)和仅BH 3调节蛋白(例如,出价)。尽管最近在表征方面取得了进展,
由于Bcl-2蛋白质的结构,该领域缺乏对蛋白质-蛋白质和蛋白质-脂质的机制的理解,
介导MOMP的相互作用。这些知识对于为未来奠定基础至关重要
开发旨在抑制或激活细胞凋亡的治疗策略。拟议
该项目旨在破译Bax/Bid/Bcl-xL的膜插入和重折叠途径
调节三联体和表征其膜调节的相互作用的框架内,
Bcl-2蛋白调控凋亡的嵌入式模型。我们将借鉴我们的经验,
其他膜插入蛋白,包括白喉毒素易位结构域,其具有结构
与Bcl-2蛋白相似。位点特异性标记与一组荧光(包括
各种类型的稳态和寿命淬灭和FRET)和电子顺磁共振
方法(DEER,O2/NiEDDA可访问性),辅以分子动力学计算机
模拟,将用于获得必要的结构,动态和热力学信息,
破译生理功能的机制。我们的初步数据表明,
开关和激活Bid/Bax/Bcl-xL调节三联体由静电和机械调节
脂质双层的性质。我们将追求以下具体目标:(1)表征膜-
在抗凋亡Bcl-xL中依赖性构象转换,(3)表征脂质依赖性膜
Bid/Bax/Bcl-xL调节三联体的募集,(3)表征孔-
形成Bax并被Bcl-xL破坏。通过对蛋白质分子机制的新认识-
Bax/Bcl-xL/Bid调节三联体中蛋白质和蛋白质-脂质相互作用,我们期望提供更清晰的
控制MOMP的分子途径图。此外,本研究还将揭示
膜上的蛋白质构象转换将告知生物医学的其他调节系统,
重要性
英文摘要
Summary
Apoptosis is crucial for proper development and function of cell populations in tissues, and its dys-
regulation is of major relevance for degenerative diseases and cancer. The critical step in triggering
apoptosis is the permeabilization of the mitochondrial outer membrane (MOMP). This process is tightly
regulated by the Bcl-2 family of proteins, which is subdivided into pro-apoptotic (e.g., Bax), anti-apoptotic
(e.g., Bcl-xL), and BH3-only regulator proteins (e.g., Bid). Despite recent advances in the characterization
of Bcl-2 proteins, the field lacks a mechanistic understanding of the protein–protein and protein–lipid
interactions that mediate MOMP. Such knowledge would be essential for setting the stage for the future
development of therapeutic strategies aimed at either suppressing or activating apoptosis. The proposed
project is aimed at deciphering the pathways of membrane insertion and refolding of the Bax/Bid/Bcl-xL
regulatory triad and characterizing their membrane-modulated interactions within the framework of the
Embedded Together model of apoptotic regulation by Bcl-2 proteins. We will draw from our experience with
other membrane-inserting proteins, including the diphtheria toxin translocation domain, which has structural
similarities to Bcl-2 proteins. Site-specific labeling in combination with a battery of fluorescence (including
various types of steady-state and lifetime quenching and FRET) and electron paramagnetic resonance
approaches (DEER, O2/NiEDDA accessibility), complemented by Molecular Dynamics computer
simulations, will be utilized to obtain structural, dynamic and thermodynamic information necessary for
deciphering the mechanism of physiological function. Our preliminary data indicate that conformational
switching and activation Bid/Bax/Bcl-xL regulatory triad is modulated by the electrostatic and mechanical
properties of the lipid bilayer. We will pursue the following specific aims: (1) Characterize membrane-
dependent conformational switching in anti-apoptotic Bcl-xL, (3) Characterize lipid-dependent membrane
recruitment of the Bid/Bax/Bcl-xL regulatory triad, (3) Characterize the membrane insertion pathway of pore-
forming Bax and its disruption by Bcl-xL. By gaining new insights into molecular mechanisms of protein–
protein and protein–lipid interactions in the Bax/Bcl-xL/Bid regulatory triad, we expect to provide a clearer
map of the molecular pathways controlling MOMP. In addition, this study will reveal general principles of
protein conformational switching on membranes that will inform other regulatory systems of biomedical
importance.
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Location of TEMPO-PC in Lipid Bilayers: Implications for Fluorescence Quenching.
TEMPO-PC 在脂质双层中的位置:对荧光猝灭的影响。
DOI:
10.1007/s00232-019-00094-1
发表时间:
2020
期刊:
The Journal of membrane biology
影响因子:
--
作者:
[Kyrychenko,Alexander, Ladokhin,AlexeyS]
通讯作者:
Ladokhin,AlexeyS
DOI:
10.1016/bs.mie.2020.12.016
发表时间:
2021
期刊:
Methods in enzymology
影响因子:
--
作者:
[Ladokhin AS, Kyrychenko A, Rodnin MV, Vasquez-Montes V]
通讯作者:
Vasquez-Montes V
DOI:
10.3390/ijms22179388
发表时间:
2021-08-30
期刊:
International journal of molecular sciences
影响因子:
5.6
作者:
[Tyagi V, Vasquez-Montes V, Freites JA, Kyrychenko A, Tobias DJ, Ladokhin AS]
通讯作者:
Ladokhin AS
Lipid-modulation of membrane insertion and refolding of the apoptotic inhibitor Bcl-xL.
细胞凋亡抑制剂 Bcl-xL 的膜插入和重折叠的脂质调节。
DOI:
10.1016/j.bbapap.2019.04.006
发表时间:
2019
期刊:
Biochimica et biophysica acta. Proteins and proteomics
影响因子:
--
作者:
[Vasquez-Montes,Victor, Vargas-Uribe,Mauricio, Pandey,NitinK, Rodnin,MykolaV, Langen,Ralf, Ladokhin,AlexeyS]
通讯作者:
Ladokhin,AlexeyS
DOI:
10.1002/pro.4742
发表时间:
2023-09
期刊:
Protein science : a publication of the Protein Society
影响因子:
--
作者:
[]
通讯作者:
Elucidating the Molecular Mechanisms of Conformational Switching during Protein Insertion into Membranes
-
批准号:10737458
-
项目类别:
-
资助金额:$60.19万
-
财政年份:2023
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:7931127
-
项目类别:
-
资助金额:$9.4万
-
财政年份:2009
-
负责人:ALEXEY LADOKHIN
-
依托单位:
EFFECT OF HEMIFLUORINATED SURFACTANTS ON MEMBRANE INSERTION/FOLDING OF DIPHTHER
-
批准号:7381284
-
项目类别:
-
资助金额:$7.39万
-
财政年份:2006
-
负责人:ALEXEY LADOKHIN
-
依托单位:
EFFECT OF HEMIFLUORINATED SURFACTANTS ON MEMBRANE INSERTION/FOLDING OF DIPHTHERI
-
批准号:7170527
-
项目类别:
-
资助金额:$7.29万
-
财政年份:2005
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:7118725
-
项目类别:
-
资助金额:$14.35万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:6709436
-
项目类别:
-
资助金额:$22.02万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:7265331
-
项目类别:
-
资助金额:$13.94万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:7470181
-
项目类别:
-
资助金额:$11.76万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:6930643
-
项目类别:
-
资助金额:$14.7万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
pH-Triggered Membrane Insertion of Proteins
-
批准号:7475811
-
项目类别:
-
资助金额:$25.7万
-
财政年份:2004
-
负责人:ALEXEY LADOKHIN
-
依托单位:
FLUORESCENCE LIFETIME STUDIES IN BIOLOGICAL MEMBRANES
-
批准号:5222613
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:ALEXEY LADOKHIN
-
依托单位:--
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