Structural studies of gasdermin E and its recognition by caspase-3.
Gasdermin E 的结构研究及其被 caspase-3 识别。
基本信息
- 批准号:10571048
- 负责人:
- 金额:$ 8.05万
- 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:2022-11-03 至 2024-10-31
- 项目状态:已结题
- 来源:
- 关键词:AddressAdoptedApoptosisApoptoticAutoimmune DiseasesBacterial InfectionsBindingBiochemicalBiological AssayCASP1 geneCASP3 geneCaspaseCell DeathCell membraneCellular biologyComplexCrohn&aposs diseaseCrystallizationCrystallographyDataDefense MechanismsDevelopmentE proteinEpithelial CellsExtravasationFamilyFamily memberHost DefenseHumanImmuneInflammasomeInflammatoryInjury to KidneyKnowledgeLengthLiposomesLyticMediatingMembraneMolecularMolecular ConformationMucositisMusMutagenesisPathogenesisPeptide HydrolasesPeptidesPlayReagentReportingRestRoleSignal PathwaySiteStructureTechniquesTestingVirus DiseasesX ray diffraction analysisX-Ray Crystallographycytokineenzyme substrate complexinhibitorinsightintestinal epitheliumkidney fibrosismembermembrane assemblymicrobialpreventrecruitsuccesstissue injuryuptake
项目摘要
Abstract
The canonical and noncanonical inflammasome signaling pathways can induce pyroptosis and secretion of
inflammatory cytokines, which function as crucial immune defense mechanisms in microbial killing and
clearance. Pyroptosis is a type of regulated lytic cell death that is mediated by members of the gasdermin family
that assemble membrane pores upon cleavage by proteases. Gasdermin E (GSDME) can be cleaved by
apoptotic caspases-3 and 7, which then triggers plasma membrane pore formation and pyroptosis instead of
apoptosis. GSDME-mediated pyroptosis and cytokine release play crucial roles in host defense against viral or
bacterial infections. On the other hand, GSDME in intestinal epithelial cells has been implicated in mucosal
inflammation and pathogenesis of Crohn’s disease, and GSDME-mediated pyroptosis contributes to renal
fibrosis and kidney injury. Despite recent progress in our understanding of the recruitment and recognition of
gasdermin D (GSDMD) by inflammatory caspases, the molecular mechanisms of GSDME recognition and
cleavage by apoptotic caspases have been poorly defined. This proposal aims to address the critical gaps in our
knowledge using complementary structure-function approaches. We hypothesize that inactive GSDME is
maintained in an autoinhibited conformation through intramolecular amino-terminal domain (NTD)-carboxy-
terminal domain (CTD) interactions, which is recognized by apoptotic caspase-3 that cleaves at the NTD-CTD
linker region to release the autoinhibition. This study will explore the structural mechanisms of GSDME
autoinhibition and its recognition by caspase-3 to test the above hypothesis. The following specific aims are built
on our past and ongoing studies on gasdermins and pyroptosis. Aim 1 will characterize the molecular
mechanisms of GSDME autoinhibition. Our preliminary data show that GSDME protein can be crystallized and
the crystals diffracted X-ray well. We will determine the structure of GSDME at its autoinhibitory state using
crystallographic approaches, and validate the NTD-CTD interface through mutagenesis studies of the interface
residues using LDH release, PI uptake, and liposome leakage assays. Aim 2 will define the mechanisms of
GSDME recognition by caspase-3. We will probe the interaction between GSDME and caspase-3 through
biochemical and cell biology techniques, and determine the structure of GSDME in complex with caspase-3
using X-ray crystallography. The success of this proposal will not only reveal important insights into GSDME
autoinhibition and recognition by apoptotic caspases that move the field forward, but also facilitate the
development of specific inhibitors for apoptotic caspase-3 based on the distinct GSDME-caspase-3 interface,
which may be valuable reagents in the study of both pyroptosis and apoptosis signaling pathways.
摘要
经典和非经典炎性体信号通路可诱导细胞火灾和分泌
炎性细胞因子,其在微生物杀伤中作为重要的免疫防御机制起作用,
间隙细胞凋亡是一种由gasdermin家族成员介导的调节性溶解性细胞死亡
其在被蛋白酶切割时组装膜孔。Gasdermin E(GSDME)可以通过以下方式裂解:
凋亡的半胱天冬酶3和7,然后触发质膜孔形成和细胞凋亡,而不是
凋亡GSDME介导的细胞凋亡和细胞因子释放在宿主防御病毒或
细菌感染另一方面,肠上皮细胞中的GSDME与粘膜有关
克罗恩病的炎症和发病机制,GSDME介导的焦亡有助于肾
纤维化和肾损伤。尽管我们最近在了解招募和承认
Gasdermin D(GSDMD)通过炎症性半胱天冬酶,GSDME识别的分子机制,
细胞凋亡半胱天冬酶的裂解还不清楚。该提案旨在解决我们在以下方面的关键差距:
使用互补结构-功能方法的知识。我们假设不活跃的GSDME是
通过分子内氨基末端结构域(NTD)-羧基-
末端结构域(CTD)相互作用,由在NTD-CTD处裂解的凋亡caspase-3识别
连接子区域以释放自身抑制。本研究将探讨GSDME的结构机制
自抑制及其被半胱天冬酶-3识别以检验上述假设。具体目标如下:
我们过去和正在进行的关于gasdermins和pyroptosis的研究目标1将表征分子
GSDME自抑制机制。我们的初步数据表明,GSDME蛋白可以结晶,
晶体很好地衍射X射线。我们将确定GSDME在其自抑制状态下的结构,
晶体学方法,并通过界面的诱变研究验证NTD-CTD界面
使用LDH释放、PI摄取和脂质体渗漏测定法测定残留物。目标2将定义
通过caspase-3识别GSDME。我们将探讨GSDME和caspase-3之间的相互作用,
生物化学和细胞生物学技术,并确定GSDME与caspase-3复合物的结构
使用X射线晶体学。这一提案的成功不仅将揭示对GSDME的重要见解,
凋亡半胱天冬酶的自身抑制和识别,使该领域向前发展,但也促进了细胞凋亡。
基于不同的GSDME-caspase-3界面开发凋亡caspase-3的特异性抑制剂,
这可能是研究细胞凋亡和细胞凋亡信号通路的有价值的试剂。
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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Tsan Sam Xiao其他文献
132 : Sensing of DNA by innate immune receptors
- DOI:
10.1016/j.cyto.2013.06.135 - 发表时间:
2013-09-01 - 期刊:
- 影响因子:
- 作者:
Tengchuan Jin;Jiansheng Jiang;Patrick Smith;Tsan Sam Xiao - 通讯作者:
Tsan Sam Xiao
Tsan Sam Xiao的其他文献
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{{ truncateString('Tsan Sam Xiao', 18)}}的其他基金
Molecular mechanisms of gasdermin recognition by proteases and autophagy proteins in cytokine release
细胞因子释放中蛋白酶和自噬蛋白识别gasdermin的分子机制
- 批准号:
10024454 - 财政年份:2020
- 资助金额:
$ 8.05万 - 项目类别:
Molecular mechanisms of gasdermin recognition by proteases and autophagy proteins in cytokine release
细胞因子释放中蛋白酶和自噬蛋白识别gasdermin的分子机制
- 批准号:
10654580 - 财政年份:2020
- 资助金额:
$ 8.05万 - 项目类别:
Molecular mechanisms of gasdermin recognition by proteases and autophagy proteins in cytokine release
细胞因子释放中蛋白酶和自噬蛋白识别gasdermin的分子机制
- 批准号:
10441356 - 财政年份:2020
- 资助金额:
$ 8.05万 - 项目类别:
Molecular mechanisms of gasdermin recognition by proteases and autophagy proteins in cytokine release
细胞因子释放中蛋白酶和自噬蛋白识别gasdermin的分子机制
- 批准号:
10223159 - 财政年份:2020
- 资助金额:
$ 8.05万 - 项目类别:
Mechanisms of tunable posttranslational control of T-cell homeostasis and tolerance
T 细胞稳态和耐受性的可调节翻译后控制机制
- 批准号:
10410503 - 财政年份:2019
- 资助金额:
$ 8.05万 - 项目类别:
Mechanisms of tunable posttranslational control of T-cell homeostasis and tolerance
T 细胞稳态和耐受性的可调节翻译后控制机制
- 批准号:
10631929 - 财政年份:2019
- 资助金额:
$ 8.05万 - 项目类别:
Molecular mechanisms of gasdermins and pyroptosis
Gasdermin 和细胞焦亡的分子机制
- 批准号:
10112920 - 财政年份:2018
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Developing chemical probes that target specific inflammasomes
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9101571 - 财政年份:2016
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$ 8.05万 - 项目类别:
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