CYTOCHROME C OXIDASE IN HEALTH AND DISEASE

细胞色素C氧化酶在健康和疾病中的作用

• 批准号: 7756627
• 负责人: Antoni Barrientos
• 金额: $ 22.06万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7756627
• 关键词: Affinity Chromatography; Binding Proteins; Biochemical Genetics; Biogenesis; Biological Assay; Catalytic Domain; Cells; Co-Immunoprecipitations; Complex; Cytochrome-c Oxidase Deficiency; Defect; Disease; Down-Regulation; Encephalopathies; Enzymes; Event; Genetic; Health; Homologous Gene; Human; Hybrids; Leigh Disease; Mass Spectrum Analysis; Membrane; Messenger RNA; Metabolism; Methods; Mitochondria; Modeling; Molecular; Molecular Weight; Muscle Weakness; Mutation; Nature; Nuclear; Oxidases; PTGS1 gene; Patients; Principal Investigator; Process; Proteins; Regulation; Reporting; Research; Respiration; Role; Source; System; Techniques; Testing; Time; Translations; Yeasts; base; cell type; clinical phenotype; crosslink; cytochrome c oxidase; mutant; polypeptide; programs; temperature sensitive mutant

Cytochrome c oxidase (COX) deficiency is the most frequent cause of mitochondrial neuromyopathiesin humans. Patients afflicted with these diseases present heterogeneous clinical phenotypes, including Leigh syndrome, muscle weakness and encephalomyopathy.A complete understanding of COX biogenesis is essential for elucidating the molecular basis underlying this group of diseases. The main objective of the proposed research is to use the yeast Saccharomycescerevisiae as a model to investigate COXassembly in wild type cells and in cells with mutations in evolutionary conservedassembly factors. Several specific aims will be pursued. 1) We have recently reported that Shylp, the yeast homologue of humanSurMp, responsible for most cases of Leigh's syndrome,catalyzes the formation of a COX assembly intermediate involving Coxlp, a mitochondrially encoded catalytic subunit of COX. The role of Shylp in expression of Coxlp will be studied. 2) More recent evidence indicates that this intermediate regulates Cox1 pexpression in a process involving other COX metabolism factors, such as Mss51p and Cox14p.The mechanisms by which these proteins regulate COX expression will be studied. Appropriately tagged Mss51p and Cox14p will be purified from over-expressing yeast cells. The availability of purified proteins will permit hypotheses concerning their activities to be tested directly. 3) The proteins involved in regulation of Coxlp synthesis by COX assembly are likely to interact transiently or permanently amongthem to perform their functions. The nature of these interactions will be characterized. In summary,the yeast system will be explored as a means of deciphering the general principles operating in the assembly of a complex membrane enzyme composed of subunit polypeptides derived from two spatially separated genetic sources. The yeast paradigm will be exploited by biochemical and genetic meansto gain a complete understanding of the function of Shylp and therefore of Surflp as well, and to clarifythe molecular basis of human COX deficiencies.

细胞色素 C 氧化酶 (COX) 缺乏是线粒体神经肌病的最常见原因 人类。患有这些疾病的患者呈现出异质的临床表型，包括 Leigh 综合征、肌无力和脑肌病。对 COX 生物发生的完整了解是 对于阐明这组疾病的分子基础至关重要。该计划的主要目标 拟议的研究是使用酵母酿酒酵母作为模型来研究 COX 组装 在野生型细胞和进化保守组装因子发生突变的细胞中。具体几个 将追求目标。 1) 我们最近报道了Shylp，人类SurMp的酵母同源物， 负责大多数 Leigh 综合征病例，催化 COX 组装中间体的形成 涉及 Coxlp，COX 的线粒体编码催化亚基。 Sylp 在表达中的作用 Coxlp将被研究。 2) 最近的证据表明该中间体调节 Cox1 p 表达 在涉及其他 COX 代谢因子（例如 Mss51p 和 Cox14p）的过程中。其机制是 这些蛋白调节COX表达的方式将被研究。适当标记的 Mss51p 和 Cox14p 将 从过度表达的酵母细胞中纯化。纯化蛋白质的可用性将允许假设 关于他们的活动要直接进行测试。 3) 参与调节Coxlp合成的蛋白质 COX 组件可能会在它们之间短暂或永久地相互作用以执行其功能。这 这些相互作用的性质将得到表征。综上所述，酵母系统将作为一种手段进行探索 破译复杂膜酶组装的一般原理 源自两个空间上分离的遗传来源的亚基多肽。酵母范式将是 通过生化和遗传学手段充分了解 Sylp 和 因此 Surflp 也是如此，并阐明人类 COX 缺陷的分子基础。