Structural and functional characterization of the human brain D-aspartate oxidase.
Structural and functional characterization of the human brain D-aspartate oxidase.
复制标题
人脑 D-天冬氨酸氧化酶的结构和功能特征。
DOI：
10.1093/oxfordjournals.jbchem.a021655
复制
发表时间：
1997
影响因子：
2.7
通讯作者：
R. Miura
中科院分区：
文献类型：
--
作者：
C. Setoyama;R. Miura
用RT-PCR方法从人脑RNA中提取D-天冬氨酸氧化酶（DDO）cDNA。DDO-1和DDO-2两种形式的mRNA被检测到。DDO cDNA和基因组DNA的结构分析表明，DDO-1和DDO-2是由单个基因的选择性剪接产生的。由DDO-1 cDNA编码的蛋白质由341个氨基酸组成，并且DDO-2的氨基酸序列与DDO-1的氨基酸序列相同，除了缺失覆盖DDO-1的残基95-153的59个氨基酸。使用大肠杆菌中的表达系统获得DDO-1的均质制剂。DDO-1选择性催化D-天冬氨酸及其N-甲基化衍生物N-甲基D-天冬氨酸的氧化脱氨; D-天冬氨酸的K（m）和k（cat）值为2.7 mM和52.5 mol D-天冬氨酸氧化物x s（-1）x mol（-1），N-甲基D-天冬氨酸的K（m）和k（cat）值为6.8 mM和37.7 mol N-甲基D-天冬氨酸氧化物x s（-1）x mol（-1），分别
D-Aspartate oxidase (DDO) cDNAs were isolated from the human brain RNA using the RT-PCR method. Two forms (DDO-1 and DDO-2) of DDO mRNA were detected. Structural analysis of the DDO cDNAs and genomic DNA showed that DDO-1 and DDO-2 are produced by alternative splicing from a single gene. A protein encoded by the DDO-1 cDNA consists of 341 amino acids, and the amino acid sequence of DDO-2 was identical to that of DDO-1 except for the absence of 59 amino acids covering residues 95-153 of DDO-1. A homogenous preparation of DDO-1 was obtained using an expression system in Escherichia coli. DDO-1 selectively catalyzed the oxidative deamination of D-aspartate and its N-methylated derivative, N-methyl D-aspartate; the values of K(m) and k(cat) for D-aspartate were 2.7 mM and 52.5 mol D-aspartate oxidized x s(-1) x mol(-1) and those for N-methyl D-aspartate were 6.8 mM and 37.7 mol N-methyl D-aspartate oxidized x s(-1) x mol(-1), respectively.