Characterizing tafazzin and Barth syndrome mutant tafazzins

表征tafazzin和巴斯综合征突变体tafazzin

基本信息

批准号：
7475127
负责人：
Steven Michael Claypool
金额：
$ 9万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2008-09-24
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7475127
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 Cartoons Cell Culture System Cellular biology Classification Complex Condition Cultured Cells Cyclic Neutropenia Cytosol Data Defect Diabetes Mellitus Disease Dissection Electrophoresis Enzyme-Linked Immunosorbent Assay Fatty Acids Fatty acid glycerol esters Figs - dietary Future General Population Goals Growth Heart Diseases Human Hyperthyroidism Individual Injury Inner mitochondrial membrane Institutes Knowledge Lecithin Ligands Light Link Lipids Localized 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 Range Reagent Recombinants Relative (related person)Research Research Personnel Role Saccharomyces cerevisiae Secure Series Skeletal system Specificity Standards of Weights and Measures System Testing Therapeutic Variant Work Yeast Model System Yeasts base boys diabetic cardiomyopathy insight interfacial loss of function mitochondrial membrane monolysocardiolipin mutant novel polyacrylamide progesterone 11-hemisuccinate-(2-iodohistamine)programs

项目摘要

DESCRIPTION (provided by applicant): 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 mitochondrial 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 cataboIism. 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 Taz1p, 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 Taz1p 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.

描述（由申请人提供）：为了开始解剖心磷脂（CL）重塑的过程，该建议的重点是推定的CL酰基转移酶Tafazzin（TAZLP）的表征，Tafazzin（TAZLP）是与Barth综合征（BTHS）相关的突变基因产物。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，哪些步骤受到调节以及正常和异常CL重塑相对于线粒体功能的后果。这将提供一个理解TAZ1 P在BTH中的作用的基础。此外，这项工作将阐明其他心肌病，包括糖尿病心肌病，最近已证明涉及CL合成的缺陷和CL Calsaboiism的增加。第一个特定目的的目的是鉴定脂质和蛋白质，这些脂质和蛋白质在复合物中直接相互作用或与TAZ1 P相互作用。初步结果表明，TAZ1 P与磷脂和/或脂肪酰基链以及组装成几种大型的大分子复合物。为了鉴定潜在的脂质配体，将采用一系列生化和遗传方法，包括使用重组TAZ1P，定义的磷脂，ELISA，FAT BLOTS，BN-PAGE，BN-PAGE和双重亲和力标记为TAZ1 P结构。双重亲和力标记的TAZ1 P结构还将通过标准生化程序揭示相互作用的蛋白质。预计该目标的结果将洞悉TAZ1P参与的CL重塑途径，包括潜在地识别TAZ1 P的底物和目标特异性，参与此过程的其他组件，以及TAZ1 P的潜在新功能。对于第二个特定目的，将研究一组BTHS突变体，该突变体发生在人和酵母直系同源物之间的相同或保守的残基上，以正确地定位在线粒体上和内部，与蛋白质和/或脂质相关，并与AIM 1中鉴定出的蛋白质和/或脂质相关联。通过与野生型TAZ1 P进行系统的比较，将提供TAZ1 P突变体和BTH的分子基础。对于那些在人类TAZ1 P独有的残基上发生的BTHS突变，将开发出细胞培养模型系统，该系统将允许该子集的BTHS突变体的分子解剖。特定目的3的目的。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，这项研究将洞悉导致心脏病的基本机制。