Structure-Function Analysis of Pro-Apoptotic BMRP and Its Role as a Tumor Suppressor

促凋亡 BMRP 的结构功能分析及其作为肿瘤抑制因子的作用

• 批准号: 10204037
• 负责人: Maribel Gonzalez-Garcia
• 金额: $ 10.35万
• 依托单位: TEXAS A&M UNIVERSITY-KINGSVILLE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204037
• 关键词: Adult; Alanine; Alzheimer' s Disease; Amino Acids; Apoptosis; Apoptotic; Area; Automobile Driving; BCL-2 Protein; BCL2 gene; Bibliography; Binding; Binding Proteins; Biochemical; Biological Assay; Cancer cell line; Cell Death; Cell physiology; Chimeric Proteins; Co-Immunoprecipitations; Collaborations; Dementia; Development; Disease; Etiology; Future; Genes; Goals; Healthy People 2020; Heart Arrest; Heart Diseases; Homeostasis; Huntington Disease; Immune System Diseases; Impairment; Infertility; Innovative Therapy; Investigation; Knowledge; Life; Maintenance; Malignant Neoplasms; Mediating; Messenger RNA; Mission; Mitochondria; Molecular; Molecular Mechanisms of Action; Molecular Target; Mutagenesis; Neurodegenerative Disorders; Organism; Pathogenesis; Peptides; Play; Prevention; Proteins; Public Health; Recording of previous events; Regulation; Reporting; Research; Resistance; Ribosomal Proteins; Role; Science; Signal Transduction; Stroke; Structure; Testing; Therapeutic Agents; Tissues; Tumor Suppressor Proteins; United States National Institutes of Health; United States Public Health Service; Yeasts; burden of illness; cancer prevention; chemoradiation; combinatorial; design; disability; falls; genetic analysis; genetic regulatory protein; human disease; innovation; insight; lung cancer cell; malignant breast neoplasm; mimetics; mutant; novel; novel therapeutic intervention; novel therapeutics; prevent; pro-apoptotic protein; protein expression; protein function; tool; tumor; yeast two hybrid system

Abstract Cell death by apoptosis is required for the normal development of metazoans, as well as for the maintenance of tissue homeostasis and normal functioning of the adult organism. Abnormal levels of apoptosis have been implicated in the etiology of human diseases including cancer, disorders of the immune system, infertility, neurodegenerative diseases, and cardiac arrest and stroke damage. In the case of cancer, impaired apoptosis is a major cause of resistance to chemo- and radio-therapies. Bcl-2 is an anti- apoptotic protein that plays a major regulatory role in this process of cell demise and, despite intense investigation, several aspects of its mechanisms of action and regulation are still not well understood. To gain further insight into the molecular mechanisms that mediate Bcl-2's anti-apoptotic function and its regulation, we conducted a search of Bcl-2 interacting proteins, which identified BMRP as a novel Bcl-2 binding protein. Current knowledge suggests that BMRP is a pro-apoptotic protein that plays a tumor- suppressor role. Our long-term goal is to enhance our understanding of the molecular mechanisms that mediate Bcl-2's pro-survival activity and its regulation, and to exploit this knowledge for the development of novel therapeutic approaches. Our hypothesis is that BMRP is a relevant component of the molecular apoptotic machinery, that its pro-apoptotic activity counteracts the activity of Bcl-2, and that mechanistic studies will yield novel targets and tools for modulating apoptosis. An increased knowledge of the molecular mechanistic basis of BMRP and Bcl-2 function and modulation will enhance our understanding of the players and signaling in apoptosis pathways, which will contribute to the development of novel drugs to prevent and/or treat human diseases involving deregulated apoptosis. Two specific aims are proposed to test our hypothesis. In the first aim, chimeric and alanine substitution mutants of BMRP will be generated to identify a killer peptide (or peptides) for future development of novel tumor combinatorial therapies, as well as amino acid residues essential to its pro-apoptotic activity. Additionally, the ability of these mutants to bind to Bcl-2 will be tested by yeast Two-Hybrid and co-immunoprecipitation assays. A second aim will investigate the role of BMRP as a tumor suppressor by determining the expression levels of the BMRP protein and mRNA in cancer cell lines, and analyzing the genetic mechanisms responsible for abnormal BMRP expression in these cancer cell lines. The proposed research is innovative, as it will establish unknown molecular aspects of BMRP function and regulation, and BMRP's role as a tumor suppressor. This knowledge is relevant to public health, since it should provide novel targets and tools for the design of advanced therapies for the treatment and/or prevention of human diseases characterized by deregulated apoptosis, such as cancer.

抽象的 细胞凋亡导致的死亡是后生动物正常发育所必需的，也是 维持组织稳态和成人有机体的正常功能。异常水平 细胞凋亡与人类疾病的病因学有关，包括癌症、 免疫系统、不孕不育、神经退行性疾病、心脏骤停和中风损伤。在这种情况下 对于癌症来说，细胞凋亡受损是对化疗和放疗产生耐药性的主要原因。 Bcl-2 是一种抗 凋亡蛋白在细胞死亡的过程中发挥着重要的调节作用，尽管存在强烈的 调查显示，其作用机制和调节的几个方面仍不清楚。到 进一步了解介导 Bcl-2 抗凋亡功能的分子机制及其作用 调节，我们对 Bcl-2 相互作用蛋白进行了搜索，将 BMRP 确定为一种新型 Bcl-2 结合蛋白。目前的知识表明 BMRP 是一种促凋亡蛋白，在肿瘤中发挥作用 抑制器的作用。我们的长期目标是增强我们对分子机制的理解 介导 Bcl-2 的促生存活动及其调节，并利用这些知识来开发 新的治疗方法。我们的假设是 BMRP 是分子的相关组成部分 凋亡机制，其促凋亡活性抵消了 Bcl-2 的活性，并且该机制 研究将产生调节细胞凋亡的新靶点和工具。增加分子知识 BMRP 和 Bcl-2 功能和调节的机制基础将增强我们对 细胞凋亡途径中的参与者和信号传导，这将有助于新药物的开发 预防和/或治疗涉及细胞凋亡失调的人类疾病。提出了两个具体目标 检验我们的假设。在第一个目标中，将产生 BMRP 的嵌合和丙氨酸取代突变体 确定一种或多种杀伤肽，用于未来开发新型肿瘤组合疗法 作为其促凋亡活性所必需的氨基酸残基。此外，这些突变体结合的能力 Bcl-2 将通过酵母双杂交和免疫共沉淀测定进行测试。第二个目标将 通过测定 BMRP 的表达水平来研究 BMRP 作为肿瘤抑制因子的作用 癌细胞系中的蛋白质和mRNA，并分析导致异常的遗传机制 这些癌细胞系中的 BMRP 表达。拟议的研究具有创新性，因为它将建立 BMRP 功能和调节的未知分子方面，以及 BMRP 作为肿瘤抑制因子的作用。这 知识与公共卫生相关，因为它应该为设计提供新颖的目标和工具 用于治疗和/或预防以放松管制为特征的人类疾病的先进疗法 细胞凋亡，例如癌症。

项目成果

A simple drying solution that minimizes cracking during air-drying of polyacrylamide gels.

一种简单的干燥解决方案，可最大限度地减少聚丙烯酰胺凝胶风干过程中的破裂。

• DOI: 10.2144/btn-2022-0094
• 发表时间: 2023
• 期刊: BioTechniques
• 影响因子: 2.7
• 作者: Gomez,KristanM;Patil,AbhijeetA;Reddig,WilliamJ;Alcala,JulisaM;González-García,Maribel;Pérez-Ballestero,Rafael
• 通讯作者: Pérez-Ballestero,Rafael