Regulation of Novel Mitochondrial Uncoupling Proteins

新型线粒体解偶联蛋白的调控

• 批准号: 6800843
• 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 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6800843
• 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(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型糖尿病的发生有关。这项建议的一个主要焦点是研究UCP2的功能和胰岛β细胞生物能量学，以努力澄清这种联系。我们将使用我们的UCP2酵母表达系统来进行参考实验，这是确认UCP2固有的特定属性所必需的。因此，将在体外使用重组酵母表达的UCP2和从胰腺13细胞中分离的线粒体进行实验。该提案的具体目的是：研究重组酵母表达的UCP2对底物的需求，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作用，并鉴定UCP2的其他天然和人工抑制剂。目的：研究经PPAR激动剂处理或未处理的胰岛β细胞线粒体的生物能量学。目标1和目标2将采用质子通量的重组和测量。它们还将包括一项假设的测试，即UCP2由脂肪酸诱导的质子运输显示出对辅酶Q10的绝对需求。AIM 3将利用敏感和精确的氧气消耗测量来检测Beta细胞线粒体中UCP2的活性。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生化调节改变所起的作用。