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.

细胞凋亡是一种正常的生理细胞自杀计划，在脊椎动物和无脊椎动物中高度保守。 该细胞死亡程序在正常发育和组织稳态中起着至关重要的作用，消除了有机体中不需要的细胞，包括受损和病毒感染的细胞。 凋亡反应的破坏与癌症，细胞死亡太少以及退行性疾病有关，而细胞死亡过多。一个称为caspase的半胱氨酸蛋白酶家族，与哺乳动物白介素1b转化酶（ICE/CASPASE-1）和CED-3有关，CED-3是线虫C.秀丽隐杆线虫的自杀基因的产物，在启动和下游程序中起着核心作用。来自Autographica ucitagrophica Nucleopolyhedrovirus的基因p35，一种杆状病毒，抑制了胱天蛋白酶的广泛范围，从而抑制了凋亡。 需要结构研究来解决涉及与caspase和p35裂解结合的p35抑制机制之间的关系。最近，我们确定了p35的晶体结构，这是父授予的第一个特定目的。 最近，我们在动物界发现了p35的第一个同源物Slp49，这是一种来自spodoptera littoralis nutlepolyhedrovirus（SINPV）的基因。 SLP49序列预测了49KDA多肽（446个氨基酸），其身份为48.8％，对P35（299个氨基酸）。 p35对诱变和功能分析的第一个同源物的可用性，加上p35和SLP49结构的确定能力，为这项研究提供了框架，旨在定义在p35样的凋亡抑制器（或调节剂）中保守的重要基序。这将为您铺平道路，以找出是否存在p35样基因家族在无脊椎动物和脊椎动物中是否存在，通过隔离其他杆状病毒和生物体的p35样基因，并建立靠背的靠背，以合成新型的小肽或肽样分子，具有抑制/模块化的凋亡样分子。