Analysis of Proteins that Modulate Apoptosis

调节细胞凋亡的蛋白质分析

• 批准号: 8054317
• 负责人: Maribel Gonzalez-Garcia
• 金额: $ 10.14万
• 依托单位: TEXAS A&M UNIVERSITY-KINGSVILLE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8054317
• 关键词: Alanine; Amino Acids; Apoptosis; Apoptosis Regulator; Apoptotic; Area; Autoimmune Diseases; Automobile Driving; Binding; Binding Proteins; Biological Assay; Biomedical Research; Cell Death; Cell Fractionation; Collaborations; Data; Development; Disease; Down-Regulation; Event; Future; Goals; Healthy People 2010; Heart Arrest; Heart Diseases; Knowledge; Life; Malignant Neoplasms; Mediating; Mission; Mitochondria; Molecular; Molecular Mechanisms of Action; Mutagenesis; Neurodegenerative Disorders; Outcome; Paper; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Physiology; Play; Preventive Intervention; Process; Progress Reports; Protein Analysis; Protein Overexpression; Proteins; Public Health; Publications; Recording of previous events; Regulation; Reporting; Research; Research Personnel; Ribosomal Proteins; Role; Scanning; Science; Scientist; Signal Pathway; Site-Directed Mutagenesis; Stroke; Structure; Study Section; Testing; Therapeutic Intervention; TimeLine; Training; Tumor Suppressor Proteins; Two-Hybrid System Techniques; United States National Institutes of Health; United States Public Health Service; Yeasts; base; burden of illness; cancer therapy; design; disability; falls; genetic regulatory protein; human disease; improved; innovation; insight; knock-down; meetings; mutant; next generation; novel; prevent; pro-apoptotic protein; programs; protein function; skills

DESCRIPTION (provided by applicant): Apoptosis is the morphologically defined and evolutionary conserved process by which most programmed cell deaths occur in metazoans. Cell death by apoptosis is essential to the development and normal physiology of metazoans. Deregulation of the tightly controlled apoptotic program contributes to the pathogenesis of human diseases such as cancer, neurodegenerative and autoimmune diseases, stroke, and cardiac arrest damage. The Bcl-2 protein is a powerful repressor of apoptotic cell death and, despite intense study, many questions remain regarding its molecular mechanisms of action and regulation. To gain further insight into the function and modulation of Bcl-2, a search of Bcl-2 binding proteins was performed, which resulted in the identification of BMRP as a novel Bcl-2 interacting protein. Current knowledge suggests that BMRP is an integral part of the apoptotic machinery. Our long-term goal is to enhance our understanding of the molecular mechanisms that mediate Bcl-2 anti-apoptotic activity and its regulation. Our hypothesis is that BMRP is a pro-apoptotic protein whose ability to stimulate apoptosis is regulated by binding with Bcl-2. An increased understanding of the mechanistic basis of BMRP and Bcl-2 function and modulation will enhance our knowledge of apoptotic pathways, which will facilitate the design of novel drugs to prevent and/or treat human diseases characterized by abnormal levels of apoptosis. Two specific aims are proposed to test our central hypothesis. First, deletion and alanine scanning mutagenesis will be carried out to identify domains and amino acid residues that are essential to the pro-apoptotic activity of BMRP. These studies will utilize PCR site-directed mutagenesis, followed by apoptosis assays. Additionally, the ability of these mutants to bind to Bcl-2 will be tested by yeast two-hybrid assays. Second, the fate of BMRP during apoptosis and the role that it plays in apoptosis signaling pathways will be analyzed by subcellular fractionation, and studies of the cellular effects of over-expression or down-regulation of BMRP in the absence and presence of other over-expressed apoptosis regulators. The proposed research is innovative, as it will establish unknown aspects of BMRP function and regulation, and BMRP's relationship to the anti-apoptotic activity of Bcl-2. This knowledge is relevant to public health, since it should provide novel targets for therapeutic intervention for diseases whose pathogenesis involves deregulated apoptosis, such as cancer.

描述（由申请人提供）：细胞凋亡是一个形态学定义和进化保守的过程，大多数程序性细胞死亡发生在后生动物中。细胞凋亡导致的细胞死亡对后生动物的发育和正常生理至关重要。对严格控制的凋亡程序的放松有助于癌症、神经退行性疾病和自身免疫性疾病、中风和心脏骤停损伤等人类疾病的发病机制。Bcl-2蛋白是凋亡细胞死亡的强大抑制因子，尽管研究深入，但关于其作用和调控的分子机制仍存在许多问题。为了进一步了解Bcl-2的功能和调控，研究人员进行了Bcl-2结合蛋白的搜索，结果发现BMRP是一种新的Bcl-2相互作用蛋白。目前的知识表明，BMRP是细胞凋亡机制的一个组成部分。