GLUTARIC ACIDEMIA TYPE II--MUTANT ELECTRON TRANSFER FLAVOPROTEIN

拟议研究的目的是继续研究葡萄糖 酸血症II型（GA 2），一种人类先天性氨基酸和脂肪酸缺陷 氧化，和ETF：泛醌氧化还原酶（ETF）;QO），蛋白质 在某些患者中缺乏这种药物， 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 这门学科已经持续了十多年， 学习突变如何破坏电子转移的学期目标 活性的ETF：AO，以及疾病表型（尤其是肾囊肿） 不典型增生）是由ETF：QO缺陷引起的。 在上一次拨款期间，我们 克隆了编码人和猪ETF：QO的cDNA，鉴定了9个突变体 在GA 2患者中表达的ETF：QO等位基因，在S cereviseae，结晶猪ETF：QO，并开始描绘机制 的ETF：QO氧化还原中心和泛醌之间的电子转移。 本供资期的具体目标是：（a）继续查明 ETF：导致GA 2的QO突变;（B）将这些突变与 临床表型，以及表达酶的功能异常 开发ETF：QO的详细结构-功能图;并且，使用基因 胚胎干细胞中的靶向方法，（c）开发小鼠模型， ETF：QO不足，（d）研究ETF：QO的时空表达 在发育中的老鼠 这些研究应有助于更好地了解 为什么ETF：QO缺乏症如此频繁地导致肾脏囊性发育不良， 这样做，可以更广泛地了解其他疾病的发病机制， 先天性异常也是。

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.