STRUCTURAL STUDIES OF PROTEINS THAT INHIBIT APOPTOSIS

抑制细胞凋亡的蛋白质的结构研究

• 批准号: 6053594
• 负责人: ANDREW J FISHER
• 金额: $ 3.96万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6053594
• 关键词: Baculoviridae; Caenorhabditis elegans; apoptosis; cysteine endopeptidases; enzyme activity; enzyme inhibitors; mutant; protein protein interaction; protein structure function; tissue /cell culture; virus genetics; virus protein

Apoptosis is a normal physiological cell suicide program highly conserved among vertebrates and invertebrates. This cell death program plays a critical role during normal development and tissue homeostasis, eliminating unwanted cells from the organism, including damaged and virus-infected cells. Disruptions of the apoptotic response are associated with cancer, where there is too little cell death, and with degenerative diseases, where is too much cell death. A family of cysteine proteases called caspases, related to the mammalian interleukin-1b converting enzyme (ICE/caspase-1) and to CED-3, the product of a suicidal gene from the nematode C. elegans, play a central role in the initiation and downstream execution of the apoptotic program. The gene p35 from the Autographa californica nucleopolyhedrovirus, a baculovirus, inhibits a broad spectrum of caspases, suppressing apoptosis. Structural studies are required to resolve the relationship between the inhibitory mechanism of P35 that involves binding to the caspase and P35-cleavage. Recently we have determined the crystal structure of P35 the first specific aim in the parent grant. Most recently we discovered the first homologue of p35 in the animal kingdom, slp49, a gene from the Spodoptera littoralis nucleopolyhedrovirus (SINPV). The slp49 sequence predicted a 49kDa polypeptide (446 amino acids) with 48.8 percent identity to P35 (of 299 amino acids). The availability of the first homologue of P35 for mutagenesis and functional analysis, together with the capability of determination of the structures of P35 and SLP49 provides the framework for this study that aims to define important motifs conserved in P35-like apoptotic suppressors (or modulators). This will pave the way to find out if a p35-like gene family exists in invertebrates and vertebrates through the isolation of p35-like genes from other baculoviruses and organisms, and establish the backbone to synthesize novel small peptide or peptide-like molecules with apoptotic suppressing/modulating activity.

细胞凋亡是脊椎动物和无脊椎动物中高度保守的正常生理细胞自杀程序。 这种细胞死亡程序在正常发育和组织稳态中发挥着关键作用，消除生物体中不需要的细胞，包括受损和病毒感染的细胞。 细胞凋亡反应的破坏与癌症（细胞死亡太少）和退行性疾病（细胞死亡太多）有关。半胱氨酸蛋白酶家族称为半胱天冬酶，与哺乳动物白细胞介素-1b 转换酶 (ICE/caspase-1) 和 CED-3（线虫自杀基因的产物）相关。线虫，在启动过程中发挥着核心作用。以及凋亡程序的下游执行。来自苜蓿银纹夜蛾核多角体病毒（一种杆状病毒）的 p35 基因可广泛抑制半胱天冬酶，从而抑制细胞凋亡。 需要进行结构研究来解决 P35 的抑制机制（涉及与 caspase 的结合）和 P35 裂解之间的关系。最近我们确定了P35的晶体结构，这是母基金中的第一个具体目标。 最近，我们发现了动物界中 p35 的第一个同源物 slp49，这是来自斜纹夜蛾核多角体病毒 (SINPV) 的基因。 slp49 序列预测为 49kDa 多肽（446 个氨基酸），与 P35（299 个氨基酸）具有 48.8% 的同一性。 P35 第一个同源物可用于诱变和功能分析，以及确定 P35 和 SLP49 结构的能力为本研究提供了框架，旨在定义 P35 样凋亡抑制子（或调节子）中保守的重要基序。这将为通过从其他杆状病毒和生物体中分离p35样基因来查明无脊椎动物和脊椎动物中是否存在p35样基因家族铺平道路，并建立合成具有凋亡作用的新型小肽或类肽分子的骨架。抑制/调节活动。