Characterizing tafazzin and Barth syndrome mutant tafazzins

表征tafazzin和巴斯综合征突变体tafazzin

• 批准号: 7691341
• 负责人: Steven Michael Claypool
• 金额: $ 24.9万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7691341
• 关键词: 3-Methylglutaconic aciduria type 2; Abbreviations; Acyltransferase; Affect; Affinity; Age; Aging; Amino Acids; Antibodies; Binding; Biochemical; Biochemical Genetics; Biochemistry; Biogenesis; Biological Models; Cardiac; Cardiolipins; Cardiomyopathies; Cell Culture System; Cell Culture Techniques; Cellular biology; Classification; Complex; Cyclic Neutropenia; Data; Defect; Diabetes Mellitus; Disease; Dissection; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Fatty Acids; Fatty acid glycerol esters; Felis catus; Figs - dietary; Future; General Population; Goals; Growth; Heart Diseases; Human; Hyperthyroidism; Individual; Injury; Inner mitochondrial membrane; Institutes; Knowledge; Lecithin; Ligands; Light; Link; Lipids; Macromolecular Complexes; Maintenance; Malignant Neoplasms; Mammalian Cell; Membrane; Membrane Proteins; Metabolism; Mitochondria; Mitochondrial Matrix; Modeling; Molecular; Molecular Models; Mutate; Mutation; Myopathy; Orthologous Gene; Pathology; Pathway interactions; Patients; Phosphatidyl glycerol; Phosphatidylglycerols; Phospholipids; Point Mutation; Polyacrylamide Gel Electrophoresis; Population Study; Positioning Attribute; Procedures; Process; Proteins; Puberty; Public Health; Reagent; Recombinants; Relative (related person); Research; Role; Saccharomyces cerevisiae; Secure; Series; Specificity; System; Testing; Therapeutic; Variant; Work; Yeast Model System; Yeasts; base; boys; diabetic cardiomyopathy; insight; interfacial; loss of function; mitochondrial membrane; molecular modeling; monolysocardiolipin; mutant; novel; polyacrylamide; progesterone 11-hemisuccinate-(2-iodohistamine); skeletal

To begin dissecting the processes of cardiolipin (CL) remodeling, this proposal focuses on the characterization of the putative CL acyltransferase, tafazzin (Tazlp), the mutant gene product associated with Barth syndrome (BTHS). The long-term goal of this research is to ascertain how CL is remodeled, the molecular players involved in this process, what steps are regulated, and the consequences of both normal and abnormal CL remodeling with respect to mitochondria! function. This will provide a basis to understand the role of Taz1 p in BTHS. Moreover, this work will shed light on additional cardiomyopathies, including diabetic cardiomyopathy, which has recently been shown to involve deficits in CL synthesis and increased CL cat a bo I ism. The goal of the first specific aim is to identify lipids and proteins that either directly interact or associate with Taz1 p in a complex. Preliminary results indicate that Taz1 p binds noncovalently to phospholipids and/or fatty acyl chains and assembles into several large macromolecular complexes. To identify potential lipid ligands, a battery of biochemical and genetic approaches will be employed including use of recombinant Tazlp, defined phospholipids, ELISA, Fat Blots, BN-PAGE, and a dual affinity tagged Taz1 p construct. The dual affinity tagged Taz1 p construct will additionally be employed to reveal interacting proteins by standard biochemical procedures. Results from this aim are expected to provide insight into the CL remodeling pathway in which Tazlp participates, including potentially identifying the substrate and target specificity of Taz1 p, other components involved in this process, and potentially novel functions of Taz1 p. For the second specific aim, a panel of BTHS mutants, occurring at identical or conserved residues between the human and yeast orthologs, will be investigated for their ability to localize to and within the mitochondrion correctly, associate with proteins and/or lipids as identified in aim 1, and function. Through the systematic comparison with wild type Taz1 p, the molecular basis for a Taz1 p mutant and BTHS will be provided. For those BTHS mutations that occur at residues unique to human Taz1 p, a cell culture model system will be developed that will allow the molecular dissection of this subset of BTHS mutants; the goal of specific aim 3. Results from this study are important for public health because cardiac disease affects the general population and this study will provide insight into basic mechanisms leading to cardiac disease.

为了开始剖析心磷脂（CL）重塑的过程，该提案重点是 推定的CL酰基转移酶Tafazzin（TAZLP）的表征，突变基因产物相关 与Barth综合征（BTHS）。这项研究的长期目标是确定CL如何重塑， 参与此过程的分子参与者，规范哪些步骤以及两者正常的后果 以及相对于线粒体的异常CL重塑！功能。这将提供理解的基础 TAZ1 P在BTHS中的作用。此外，这项工作将阐明其他心肌病，包括 糖尿病心肌病最近已显示出涉及CL合成的缺陷并增加 Cl Cat A Bo I ISM。第一个特定目的的目标是识别直接相互作用的脂质和蛋白质 或在复合物中与TAZ1 p相关。初步结果表明TAZ1 P与 磷脂和/或脂肪酰基链，并组装成几种大型大分子复合物。到 确定潜在的脂质配体，将采用一系列生化和遗传方法 使用重组TAZLP，定义的磷脂，ELISA，脂肪印迹，BN-PAGE和双重亲和力标记 TAZ1 P结构。双重亲和力标记的TAZ1 P结构还将用于揭示互动 蛋白质通过标准生化程序。预计该目标的结果将为您提供洞察力 TAZLP参与的CL重塑途径，包括潜在地识别底物和 TAZ1 P的目标特异性，此过程中涉及的其他组件，以及潜在的新功能 taz1 p。对于第二个特定目的，在相同或保守的残基处发生的BTHS突变体面板 在人类和酵母直系同源物之间，将研究其本地化和内部的能力 正确的线粒体，与AIM 1中鉴定的蛋白质和/或脂质相关联，并与功能相关。通过 与野生型TAZ1 P进行了系统的比较，TAZ1 P突变体的分子基础和BTH将是 假如。对于在人类TAZ1 P独有的残基上发生的BTHS突变，一种细胞培养模型 将开发系统，以使BTHS突变体的该子集的分子解剖；目标 特定目的3。这项研究的结果对公共卫生很重要，因为心脏病会影响 一般人群和这项研究将洞悉导致心脏病的基本机制。