NLRP1 and CARD8 Inflammasomes: Assembly, Regulation and Stress Sensing

NLRP1 和 CARD8 炎症小体：组装、调节和压力感应

基本信息

批准号：
10646160
负责人：
Hao Wu
金额：
$ 53.1万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646160
关键词：
Acceleration Address Architecture Attention Autoimmune Diseases Binding Biochemical Biological Biology C-terminal CASP1 gene Caspase Cell Death Cellular biology Characteristics Chemicals Color Complement Complex Cryoelectron Microscopy Data Dipeptidyl Peptidases Disease Dissociation Embryonic Development Experimental Designs Filament Germ-Line Mutation Health Homeostasis Human Immune signaling Immunity Inflammasome Inflammation Inflammatory Interleukin-1 beta Interruption Knock-out Lead Length Leucine-Rich Repeat Maintenance Mediating Modeling Molecular Molecular Profiling Mus Mutation N-terminal Nucleotides Pathogenicity Pathologic Pathway interactions Pattern recognition receptor Peptide Hydrolases Peptides Pharmaceutical Preparations Play Predisposition Process Proline Proteins Regulation Repression Rest Rodent Role Shigella Signal Transduction Signaling Protein Site Skin Specificity Stress Structure Testing VDAC1 gene anthrax lethal factor autoinflammatory autoinflammatory diseases carcinogenesis cytokine design experimental study human disease inhibitor keratinocyte marenostrin multicatalytic endopeptidase complex mutant novel pathogen recruit response sensor skin disorder small molecule inhibitor

项目摘要

Abstract Nucleotide-binding domain (NBD)-like and leucine-rich repeat (LRR)-containing proteins (NLRs) perform diverse functions in cellular biology, mediating a broad set of fundamental biological pathways from immune signaling to embryonic development. In immunity, several NLRs form supramolecular protein signaling complexes called inflammasomes. Inflammasomes activate caspase-1, an inflammatory protease that processes the cytokine interleukin-1β (IL-1β) and the pore-forming protein gasdermin D to potentiate pyroptotic cell death. One such inflammasome- forming protein, NLRP1, is directly activated in response to intracellular pathogens and the inhibition of DPP9, an endogenous peptidase, serving as both a pattern recognition receptor and a signaling complex. While most NLRs share a common domain architecture, the multifunctionality and regulation of NLRP1 requires additional structural components. In addition to the characteristic NBD and LRR domains, human NLRP1 contains an N- terminal pyrin domain (PYD) and a rare function-to-find domain (FIIND) followed by a caspase activation and recruitment domain (CARD) on its C-terminus. The only other protein with a FIIND is CARD8, which has also been shown to form inflammasomes, leading to cytokine secretion and cell death. Mechanistically, the NLRP1 and CARD8 FIIND domains constitutively catalyze autoproteolytic cleavage, leading to the formation of two noncovalently associated peptides: an autoinhibitory N-terminal fragment and an inflammatory C-terminal fragment. Additionally, Dipeptidyl peptidase 8 or 9 (DPP8/9) binds and inhibits NLRP1 and CARD8, and small molecule inhibitors of DPP8/9 induces NLRP1 and CARD8 activation through some poorly understood pathway. In this application, we propose to elucidate the activation and regulation of NLRP1 and CARD8 using a structure-guided approach. We will determine structures of NLRP1 or CARD8 in complex with DPP8 or DPP9, both WT and mutants. We will analyze the structures and perform additional biochemical and cellular biological experiments to test our hypotheses. In one central aim, we propose that NLRP1 and CARD8 are stress sensors for endogenous cellular dysregulation and play important roles in unwanted inflammation and diseases.

抽象的核苷酸结合结构域（NBD）类似和含亮氨酸的重复（LRR） - 含蛋白（NLR）在细胞生物学中执行潜水功能，介导了一系列基本生物学从免疫信号传导到胚胎发育的途径。免疫，几种NLR形式超分子蛋白信号传导复合物称为炎症。炎性症激活 caspase-1，一种处理细胞因子白介素1β（IL-1β）的炎性蛋白酶和孔形成蛋白质气体D到潜在的凋亡细胞死亡。一个这样的炎症组形成蛋白质NLRP1，响应细胞内病原体和抑制DPP9，一种内源性肽，既用作模式识别受体和信号复合物。虽然大多数NLR共享一个共同的域架构，但 NLRP1的多功能性和调节需要其他结构组件。除了特征性的NBD和LRR结构域外，人NLRP1还包含N- 末端pyrin结构域（PYD）和罕见的函数与界面结构域（fiind），然后是caspase 其C末端的激活和招聘域（CARD）。唯一的其他蛋白质 fiind是Card8，它也已显示出炎症，导致细胞因子分泌和细胞死亡。从机械上讲，NLRP1和Card8 Fiind域组成型催化催化性裂解，导致两种非共价相关的形成肽：自身抑制性N末端片段和炎症C末端片段。另外，二肽基肽酶8或9（DPP8/9）结合并抑制NLRP1和Card8，并且 DPP8/9的小分子抑制剂通过一些不良的影响NLRP1和Card8激活了解途径。在此应用中，我们建议阐明的激活和调节 NLRP1和CARD8使用结构引导的方法。我们将确定NLRP1的结构或与DPP8或DPP9复合体的Card8，无论是WT和突变体。我们将分析结构并执行其他生化和细胞生物学实验以检验我们的假设。在一个中心目的中，我们建议NLRP1和CARD8是内源性的应力传感器细胞失调并在不需要的注射和疾病中起重要作用。