The integral membrane protease ZMPSTE24, lamin A processing, and the premature aging disease progeria

整合膜蛋白酶ZMPSTE24、核纤层蛋白A加工与早衰症早衰症

• 批准号: 10207666
• 负责人: Susan D. Michaelis
• 金额: $ 45.69万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207666
• 关键词: Age; Aging; Alleles; Binding; Biochemical; Biogenesis; Biological; Biological Assay; Biology; Catalysis; Catalytic Domain; Cell Line; Cells; Child; Disease; Enzymes; Genes; Health; Human; Laboratories; Lamin Type A; Light; Longevity; Membrane; Metalloproteases; Molecular; Mutation; Nuclear Matrix; Partner in relationship; Peptide Hydrolases; Pheromone; Physiological; Play; Population; Premature aging syndrome; Prevention; Process; Progeria; Proteins; Proteolysis; Proteolytic Processing; Publishing; Quality Control; Research Project Grants; Research Subjects; Role; Series; Side; Structure; Symptoms; Syndrome; System; Water; Work; Yeasts; Zinc; healthy aging; in vivo; insight; misfolded protein; normal aging; novel; prelamin A; tool

Project Summary This project focuses on ZMPSTE24, an intriguing integral membrane zinc metalloprotease important for human health and longevity. ZMPSTE24 plays a critical role in the proteolytic processing of the farnesylated CAAX protein prelamin A, precursor of the nuclear scaffold component lamin A. Mutations in the genes encoding either prelamin A or ZMPSTE24 that block cleavage cause the severe premature aging disorder Hutchinson- Gilford Progeria syndrome (HGPS) and a set of related progeroid diseases in which an aberrant permanently farnesylated form of lamin A is the “molecular culprit” that promotes aging-related symptoms. Importantly, diminished prelamin A processing by ZMPSTE24 may also be a critical factor in normal physiological aging. My laboratory pioneered the study of ZMPSTE24 in the early years of this project. We discovered this protease in yeast, where it is called Ste24, and showed that it has two distinct proteolytic activities in the biogenesis of the mating pheromone a-factor (cleavage of the CAAX motif and a second upstream cleavage). Importantly, we demonstrated that mammalian ZMPSTE24 performs these same cleavages in prelamin A maturation. Through this sustained body of work, together with a powerful new humanized yeast system for prelamin A cleavage recently developed in my laboratory, and our recent work showing a quality control role for ZMPSTE24 in removing misfolded proteins from “clogged” translocons, we have at our fingertips a full arsenal of tools, including a variety of biochemical and in vivo assays, cell lines, and disease alleles that will facilitate the proposed studies. The recently published structure of human ZMPSTE24 and the nearly superimposable yeast Ste24 revealed surprising features, defining ZMPSTE24/Ste24p as truly novel class of intramembrane proteases. The seven transmembrane spans of ZMPSTE24/Ste24p form a voluminous water- filled intramembrane chamber with the zinc metalloprotease catalytic site facing the chamber interior, so that substrate access is restricted and must occur through one of several side portals. One major challenge in the field is to establish how this novel intramembrane protease works. We propose to mechanistically dissect human ZMPSTE24 and its substrate prelamin A to define precisely how proteolysis of prelamin A occurs inside of an intramembrane chamber. We will identify features of ZMPSTE24 and prelamin A important for substrate recognition, entry, binding, catalysis, and product release, and will determine the step at which ZMPSTE24 and lamin A disease alleles malfunction. We will also deploy powerful designer screens possible in yeast to identify new Ste24 substrates that may shed light on its mechanism. We will begin an exciting new series of studies aimed at exploring a second major challenge, namely whether and how diminished ZMPSTE24 activity and prelamin A accumulation may drive physiological aging. Together, these studies will reveal fundamental principles relevant to intramembrane protease biology, premature aging, and normal physiological aging. 1

项目摘要 该项目侧重于ZMPSTE24，这是一个有趣的整体膜锌金属蛋白酶，对人很重要 健康与寿命。 ZMPSTE24在Farnesyrated Caax的蛋白水解加工中起着至关重要的作用 蛋白质前A，核脚手架成分层状A的前体A。编码基因中的突变 阻断裂解的预胺A或ZMPSTE24会导致严重的过早衰老疾病Hutchinson- 吉尔福德富豪综合征（HGP）和一组相关的雌激素疾病，其中异常永久性 层粘连蛋白A的法尼化形式是促进与衰老相关的符号的“分子罪魁祸首”。重要的是， ZMPSTE24加工的预序列减少也可能是正常身体衰老的关键因素。 我的实验室在该项目初期对ZMPSTE24进行了启动。我们发现了这个 酵母中的蛋白酶，称为Ste24，并表明它在该酵母中具有两种不同的蛋白水解活性 交配信息素A因子的生物发生（CAAX基序的裂解和第二个上游裂解）。 重要的是，我们证明了哺乳动物ZMPSTE24在预序中执行相同的裂解 成熟。通过这种持续的工作，以及强大的新型人源酵母系统 预胺A最近在我的实验室开发的乳沟，而我们最近的工作显示了质量控制角色 对于ZMPSTE24，从“ clegged”易位的蛋白质中去除错误折叠的蛋白 工具的武器库，包括各种生化和体内测定，细胞系和疾病等位基因 促进拟议的研究。人类ZMPSTE24的最近发表的结构和几乎 可叠加的酵母ste24揭示了惊喜特征，将ZMPSTE24/Ste24p定义为真正的新颖班级 膜内蛋白酶。 ZMPSTE24/Ste24p的七个跨膜跨度形成大量的水。 填充膜内室，锌金属蛋白酶催化部位面向腔室内，以便 底物访问受到限制，必须通过几个侧门户之一进行。在 领域是建立这种新型膜内蛋白酶的工作原理。我们提议机械 剖析人ZMPSTE24及其底物预序列A，以精确定义预蛋白A的蛋白水解 发生在膜内室内。我们将识别ZMPSTE24和预胺的特征是重要的 用于底物识别，进入，绑定，催化和产品释放，并​​将确定该步骤 ZMPSTE24和椎板A疾病等位基因故障。我们还将部署强大的设计师屏幕 在酵母中，以识别可能阐明其机制的新的Ste24底物。我们将开始一个令人兴奋的新 一系列研究旨在探索第二个主要挑战，即是否以及如何减少 ZMPSTE24活性和预胺A积累可能会促进生理衰老。在一起，这些研究 将揭示与膜内蛋白酶生物学，过早衰老和正常的基本原理 生理衰老。 1