GLUTARIC ACIDEMIA -- MUTANT GLUTARYL-COA DEHYDROGENASE

拟议研究的目的是继续研究葡萄糖 酸血症I型（GAI），一种人类先天性赖氨酸和色氨酸缺陷 导致基底神经节变性和进行性神经元损伤的代谢。 儿童期运动障碍，死亡通常发生在 头十年 这种情况是由于缺乏戊二酰辅酶A 脱氢酶（GCDH），一种含有FAD的线粒体酶， 戊二酰辅酶A氧化脱羧为巴豆酰辅酶A和CO2。 我们有 一直在调查GAI和GCDH约十五年，现在 能够大大提高我们对酶如何 函数在正常状态下以及函数如何被自然扰动 突变导致GAI。 本供资期的具体目标 是（a）从猪肉和人肝中分离戊二酰辅酶A脱氢酶， 并开发氨基末端和内部肽序列;（B） 开发编码人酶的全长cDNA克隆， 通过核苷酸和氨基酸序列共线性以及 在cos细胞、成纤维细胞或蛙卵母细胞中表达;和（c）发育 通过表达正常和突变的GCDH酶， 大肠杆菌。

The objective of the proposed research is to pursue studies on glutaric acidemia type I (GAI), a human inborn error of lysine and tryptophan metabolism which causes degeneration of the basal ganglia and a progressive movement disorder in childhood, with death usually occurring during the first decade. The condition is due to deficiency of glutaryl-CoA dehydrogenase (GCDH), an FAD-containing mitochondrial enzyme that catalyzes oxidative decarboxylation of glutaryl-CoA to crotonyl-CoA and CO2. We have been investigating GAI and GCDH for some fifteen years, and are now positioned to substantially improve our understanding of how the enzyme functions in the normal state and how function is perturbed by naturally occurring mutations to cause GAI. Specific aims for this funding period are to (a) isolate glutaryl-CoA dehydrogenase from pork and human liver, and to develop amino terminal and internal peptide sequences; (b) to develop a full length cDNA clone encoding the human enzyme, authenticating its identity by colinearity of nucleotide and amino acid sequences and by expression in cos cells, fibroblasts, or frog oocytes; and (c) to develop a structure function map of GCDH by expressing normal and mutant enzymes in E coli.