GLUTARIC ACIDEMIA TYPE II--MUTANT ELECTRON TRANSFER FLAVOPROTEIN

拟议研究的目标是继续研究戊二酸。 II型酸血症(GA2)，一种人类先天的氨基酸和脂肪酸错误 氧化，以及在ETF上：泛醌氧化还原酶(ETF)；QO)，蛋白质 在一些患有这种疾病的患者中是缺乏的，而且通常 催化电子转移黄素蛋白(ETF)之间的电子转移 和线粒体呼吸链。我们对此的调查 这个课题已经持续了十多年，已经有了很长的时间 学习突变如何扰乱电子转移的学期目标 ETF的活性：AO，以及疾病的表型(尤其是肾囊性 不典型增生)是由ETF：QO缺乏引起的。在上一次授权期内，我们 克隆人和猪ETF：QO的cDNA，鉴定出9个突变体 GA2患者ETF：QO等位基因及S表达野生型和突变型ETF：QO Cereviseae，结晶猪ETF：QO，并开始描述机制 ETF：QO氧化还原中心和泛醌之间的电子转移。 本供资期间的具体目标是：(A)继续确定 ETF：导致GA2的QO突变；(B)这些突变与 临床表型和表达酶功能异常 开发ETF：QO的详细结构-功能图；并使用一个基因 在胚胎干细胞中的靶向方法，(C)建立小鼠模型 ETF：QO不足，以及(D)研究ETF：QO的时空表达 在发育中的小鼠身上。这些研究应该有助于更好地理解 为什么ETF：QO缺陷如此频繁地导致肾囊性发育不良， 通过这样做，可以更广泛地了解其他疾病的发病机制 先天性畸形也是如此。

The objective of the proposed research is to pursue studies on glutaric acidemia type II (GA2), a human inborn error of amino and fatty acid oxidation, and on ETF:ubiquinone oxidoreductase (ETF);QO), the protein which is deficient in some patients with the disease and which normally catalyzes electron transfer between electron transfer flavoprotein (ETF) and the mitochondrial respiratory chain. Our investigations on this subject, which have been ongoing for more than ten years, have the long term objective of learning how mutations disrupt the electron transfer activity of ETF:AO, and how disease phenotype (especially renal cystic dysplasia) is caused by ETF:QO deficiency. During the last grant period we cloned cDNAs encoding human and porcine ETF:QO, identified nine mutant ETF:QO alleles in GA2 patients, expressed wild type and mutant ETF:QO in S cereviseae, crystallized porcine ETF:QO, and began to delineate mechanisms of electron transfer between the ETF:QO redox centers and ubiquinone. Specific aims for this funding period are to (a) continue to identify ETF:QO mutations that cause GA2; (b) correlate these mutations with clinical phenotype, and with functional abnormalities in expressed enzyme to develop a detailed structure-function map of ETF:QO; and, using a gene targeting approach in embryonic stem cells, (c) develop a murine model of ETF:QO deficiency, and (d) study spatial and temporal expression of ETF:QO in the developing mouse. These studies should permit better understanding of why ETF:QO deficiency so frequently causes renal cystic dysplasia and, in doing so, may permit broader understanding of the pathogenesis of other congenital anomalies as well.