GLUTARIC ACIDEMIA--MUTANT GLUTARYL-COA DEHYDROGENASE

拟议研究的目标是继续研究戊二酸。 I型酸血症(GAI)--人类赖氨酸和色氨酸的先天性错误 导致基底节变性和进行性脑损伤的代谢 儿童时期的运动障碍，死亡通常发生在 第一个十年。这种情况是由于戊二酰辅酶A缺乏所致。 脱氢酶(GCDH)，一种含有FAD的线粒体酶，催化 戊二酰辅酶A氧化脱羧制巴豆酰辅酶A和二氧化碳。我们有 我调查GAI和GCDH大约15年了，现在 极大地提高了我们对酶是如何 正常状态下的功能以及功能如何被自然干扰 发生突变导致GAI。本资助期的具体目标 是：(A)从猪肉和人肝中分离出戊二酰辅酶A脱氢酶， 并开发氨基端和内部肽序列；(B) 建立编码人酶的全长cDNA克隆，鉴定 其同源性通过核苷酸和氨基酸序列的共线性以及通过 在cos细胞、成纤维细胞或青蛙卵母细胞中的表达；以及(C)发育 表达正常和突变酶的GCDH结构功能图谱 大肠埃希氏菌。

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.