Rzem Rim, Veiga-da-Cunha Maria, Noël Gaëtane, Goffette Sophie, Nassogne Marie-Cécile, Tabarki Brahim, Schöller Christina, Marquardt Thorsten, Vikkula Miikka, Van Schaftingen Emile
Laboratory of Physiological Chemistry, Christian de Duve Institute of Cellular Pathology, Université Catholique de Louvain, Avenue Hippocrate 75, B-1200 Brussels, Belgium.
Proc Natl Acad Sci U S A. 2004 Nov 30;101(48):16849-54. doi: 10.1073/pnas.0404840101. Epub 2004 Nov 17.
The purpose of this study was to identify the biochemical and genetic defect in L-2-hydroxyglutaric aciduria, a neurometabolic disorder characterized by the presence of elevated concentrations of L-2-hydroxyglutaric acid in urine, plasma, and cerebrospinal fluid. Evidence is provided for the existence in rat tissues of a FAD-dependent enzyme catalyzing specifically the oxidation of L-2-hydroxyglutarate to alpha-ketoglutarate. This enzyme is mainly expressed in liver and kidney but also at lower levels in heart, brain, and other tissues. Subcellular fractionation indicates that the liver enzyme is present in mitochondria, where it is bound to membranes. Based on this information, a database search led to the identification of a gene encoding a human hypothetical protein homologous to bacterial FAD-dependent malate dehydrogenases and targeted to mitochondria. The gene encoding this protein, present on chromosome 14q22.1, was found to be in a region homozygous in patients with L-2-hydroxyglutaric aciduria from two consanguineous families. Three mutations that replaced a highly conserved residue (Lys-71-Glu and Glu-176-Asp) or removed exon 9 were identified in homozygous state in patients from three distinct families and were found to cosegregate with the disease. It is concluded that L-2-hydroxyglutarate is normally metabolized to alpha-ketoglutarate in mammalian tissues and that L-2-hydroxyglutaric aciduria is caused by mutations in the gene that most likely encodes L-2-hydroxyglutarate dehydrogenase. The pathological findings observed in this metabolic disorder must therefore be due to a toxic effect of L-2-hydroxyglutarate on the central nervous system.
本研究的目的是确定L-2-羟基戊二酸尿症中的生化和遗传缺陷,这是一种神经代谢紊乱疾病,其特征是尿液、血浆和脑脊液中L-2-羟基戊二酸浓度升高。有证据表明,大鼠组织中存在一种依赖黄素腺嘌呤二核苷酸(FAD)的酶,该酶特异性催化L-2-羟基戊二酸氧化为α-酮戊二酸。这种酶主要在肝脏和肾脏中表达,但在心脏、大脑和其他组织中的表达水平较低。亚细胞分级分离表明,肝脏中的这种酶存在于线粒体中,并与膜结合。基于这些信息,通过数据库搜索鉴定出一个基因,该基因编码一种与细菌依赖FAD的苹果酸脱氢酶同源且定位于线粒体的人类假定蛋白。发现该蛋白的编码基因位于14号染色体q22.1区域,在来自两个近亲家庭的L-2-羟基戊二酸尿症患者中该区域为纯合状态。在来自三个不同家庭的患者中,以纯合状态鉴定出三个取代高度保守残基(赖氨酸-71-谷氨酸和谷氨酸-176-天冬氨酸)或缺失外显子9的突变,并且发现这些突变与疾病共分离。得出的结论是,L-2-羟基戊二酸在哺乳动物组织中通常代谢为α-酮戊二酸,L-2-羟基戊二酸尿症是由最有可能编码L-2-羟基戊二酸脱氢酶的基因突变引起的。因此,在这种代谢紊乱中观察到的病理发现必定是由于L-2-羟基戊二酸对中枢神经系统的毒性作用。