Cysteine Proteases in Apoptosis

半胱氨酸蛋白酶在细胞凋亡中的作用

• 批准号: 8240383
• 负责人: Emad S Alnemri
• 金额: $ 32.3万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8240383
• 关键词: Acne; Acute suppurative arthritis due to bacteria; Adaptor Signaling Protein; Affect; Apical; Apoptosis; Apoptotic; Aspartate; Biochemical; Biochemical Genetics; Biological; Caspase; Caspase-1; Cell Line; Cell physiology; Cells; Chronic; Chronic Disease; Colchicine; Complex; Cysteine Protease; Cytoskeleton; Data; Defect; Disease; Disease Pathway; Exhibits; Familial Mediterranean Fever; Family; Feedback; Genes; Genetic; Human; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-4; Lead; Mediating; Microtubules; Missense Mutation; Mus; Mutate; Mutation; Natural Immunity; Osteomyelitis; Pathway interactions; Physiological; Play; Process; Proteins; Pyoderma Gangrenosum; Research; Role; Signal Pathway; Stimulus; Symptoms; Syndrome; System; Therapeutic; abstracting; base; clinical application; cytokine; design; insight; macrophage; marenostrin; member; procaspase-1; protein complex; reconstitution; research study; response

Abstract Caspase-1 is a member of a family of aspartate-specific cystein proteases, known as the caspase family, involved in apoptosis and inflammation. Recent studies revealed that caspase-1 is activated within large protein complexes or assemblies called the inflammasomes. The inflammasomes are composed of several adaptor proteins that interact with caspase-1 and induce its oligomerization and activation. Preliminary evidence from the applicant's lab suggest that pyrin, a protein mutated in the auto- inflammatory disease Familial Mediterranean fever (FMF), plays a major role in inflammation by assembling an inflammasome complex with the adaptor protein ASC, and procaspase-1 leading to ASC oligomerization, caspase-1 activation and IL-1? processing. Pyrin also appears to play an important role in apoptosis as macrophages from pyrin-truncation mice exhibit a defect in apoptosis compared to macrophages from wildtype mice in response to IL-4 and LPS stimulation. Evidence suggests that the inflammatory and perhaps the apoptotic activities of pyrin are regulated by two cytoskeleton associated proteins known as PSTPIP1 and PSTPIP2. Missense mutations in these proteins are associated with two further auto-inflammatory diseases, known as pyogenic arthritis, pyoderma gangrenosum, acne syndrome (PAPA) and chronic multifocal osteomyelitis (cmo) syndrome, respectively. In this competing continuation application three specific aims are proposed to study the physiological role of pyrin in apoptosis and inflammation and the role of the cytoskeleton in these two pyrin pathways. In the first specific aim, the role of pyrin in caspase-1 activation will be investigated through detailed biochemical and biological studies in cell-based and cell-free reconstitution systems and human and mouse macrophage cell lines. In the second specific aim, experiments are proposed to characterize the interaction of PSTPIP1 and PSTPIP2 with pyrin and determine their ability to regulate caspase-1 activation in human and mouse cells. In the third specific aim, biological, biochemical and genetic approaches will be used to elucidate the mechanism by which pyrin regulates apoptosis. Understanding the interplay of the pyrin-mediated in?ammatory and apoptotic responses and the mechanism of activation of caspase-1 by pyrin is likely to contribute important insights into the role of pyrin in innate immunity and apoptosis, with obvious potential clinical applications. Project Narrative Mutations in the genes encoding pyrin and the pyrin-associated protein PSTPIP1 are associated with severe autoinflammatory diseases in humans. This research will investigate the physiological function of pyrin to elucidate its role in normal cellular function and how disease-associated mutations alter its function. The results of this research will help in the design and discovery of effective therapeutics to treat autoinflammatory diseases.

抽象的 caspase-1是天冬氨酸特异性西氏蛋白酶的成员，称为caspase家族， 参与凋亡和炎症。最近的研究表明，caspase-1被激活 大型蛋白质复合物或组件称为炎症。炎症组成 与caspase-1相互作用并诱导其低聚和激活的几种衔接蛋白。 申请人实验室的初步证据表明，吡啶是一种在自身中突变的蛋白 炎症性疾病家族性地中海（FMF）在炎症中起着重要作用 与衔接蛋白ASC组装炎性体复合物，并导致ASC的procaspase-1 寡聚，caspase-1激活和IL-1？加工。吡啶似乎也​​起着重要作用 在凋亡中作为pyrin截断小鼠的巨噬细胞的凋亡表现出凋亡的缺陷 响应IL-4和LPS刺激的野生型小鼠的巨噬细胞。有证据表明 炎症性以及吡啶的凋亡活性受到两个相关的细胞骨架调节 蛋白质称为PSTPIP1和PSTPIP2。这些蛋白质中的错义突变与 另外两种自动炎症性疾病，称为化脓性关节炎，脓疱性坏疽，痤疮 综合征（PAPA）和慢性多焦点骨髓炎（CMO）综合征。在这个竞争中 延续申请提出了三个具体目标来研究吡啶在 细胞凋亡和炎症以及细胞骨架在这两个吡啶途径中的作用。在第一个 具体目的，将通过详细的生化研究pyrin在caspase-1激活中的作用 以及基于细胞和无细胞的重建系统以及人类和小鼠的生物学研究 巨噬细胞系。在第二个特定目标中，提出了实验以表征 PSTPIP1和PSTPIP2与吡啶的相互作用，并确定它们调节caspase-1的能力 在人和小鼠细胞中激活。在第三个特定目标中，生物学，生化和遗传 方法将用于阐明吡啶调节凋亡的机制。理解 在弹药和凋亡反应中介导的吡啶介导的相互作用以及 pyrin激活caspase-1可能有助于对pyrin在先天物中的作用的重要见解 免疫和凋亡，具有明显的潜在临床应用。项目叙述 编码吡啶和吡啶相关蛋白PSPIP1的基因中的突变与 人类严重的自身炎症性疾病。这项研究将研究 吡啶以阐明其在正常细胞功能中的作用以及疾病相关突变如何改变其 功能。这项研究的结果将有助于设计和发现有效的治疗剂 治疗自发性疾病。