Regulation of Novel Mitochondrial Uncoupling Proteins

新型线粒体解偶联蛋白的调控

• 批准号: 6644177
• 负责人: Keith D Garlid
• 金额: $ 3.8万
• 依托单位: PORTLAND STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6644177
• 关键词: Europe; adenosine diphosphate; adenosine triphosphate; aspartate; bioenergetics; cofactor; cooperative study; diphosphonate; fatty acid transport; fungal proteins; gene expression; glutamates; hamsters; ion transport; laboratory rat; method development; miniature swine; mitochondria; noninsulin dependent diabetes mellitus; nucleoproteins; oxygen consumption; pancreatic islets; peroxisome proliferator activated receptor; phosphonate; protein structure function; recombinant proteins; yeasts

DESCRIPTION (provided by applicant) Recent evidence suggests that there is a link between enhanced expression of uncoupling protein 2 (UCP2) and development of type II diabetes. A major focus of this proposal is to investigate UCP2 function and pancreatic Beta-cell bioenergetics in an effort to clarify this connection. We will employ our yeast expression system for UCP2 to perform the reference experiments, which are required to confirm that a particular property is inherent to UCP2. Thus, experiments will be performed in vitro with recombinant yeast-expressed UCP2 and also with mitochondria isolated from pancreatic 13-cells. The Specific Aims of the proposal are: To study substrate requirements of recombinant yeast-expressed UCP2 to determine whether UCP2-mediated proton transport has a particular requirement for fatty acid chain and saturation. To study nucleotide binding and inhibition of UCP2 and to identify other natural and artificial inhibitors of UCP2. To study the bioenergetics of mitochondria isolated from pancreatic beta-cells that have been treated or untreated with PPAR agonists. Aims 1 and 2 will employ reconstitutions and measurements of proton flux. They will also include a test of the hypothesis that fatty acid-induced proton transport by UCP2 exhibits an absolute requirement for coenzyme Q10. Aim 3 will utilize sensitive and precise measurements of oxygen consumption to detect UCP2 activity in Beta-cell mitochondria. We will attempt to distinguish between the contributions of increased UCP2 expression and altered biochemical regulation of UCP2 in mitochondria from treated and untreated cells.

描述（由申请人提供） 最近的证据表明，解偶联蛋白 2 (UCP2) 的表达增强与 II 型糖尿病的发展之间存在联系。该提案的一个主要重点是研究 UCP2 功能和胰腺 Beta 细胞生物能学，以努力阐明这种联系。我们将使用我们的 UCP2 酵母表达系统进行参考实验，以确认 UCP2 固有的特定特性。因此，将使用重组酵母表达的 UCP2 以及从胰腺 13 细胞中分离的线粒体进行体外实验。该提案的具体目标是： 研究重组酵母表达的 UCP2 的底物要求，以确定 UCP2 介导的质子运输是否对脂肪酸链和饱和度有特殊要求。研究 UCP2 的核苷酸结合和抑制，并鉴定 UCP2 的其他天然和人工抑制剂。研究从经过或未经 PPAR 激动剂处理的胰腺 β 细胞中分离出的线粒体的生物能学。目标 1 和 2 将采用质子通量的重构和测量。他们还将测试以下假设：UCP2 诱导的脂肪酸质子运输对辅酶 Q10 具有绝对需求。目标 3 将利用灵敏且精确的耗氧量测量来检测 Beta 细胞线粒体中的 UCP2 活性。我们将尝试区分处理和未处理细胞线粒体中 UCP2 表达增加和 UCP2 生化调节改变的贡献。