Caenorhabditis elegans has two genes encoding functional d‐aspartate oxidases
Caenorhabditis elegans has two genes encoding functional d‐aspartate oxidases
复制标题
秀丽隐杆线虫有两个编码功能性 d-天冬氨酸氧化酶的基因
DOI：
10.1111/j.1742-4658.2006.05571.x
复制
发表时间：
2007
期刊：
影响因子：
--
通讯作者：
H. Homma
中科院分区：
文献类型：
--
作者：
M. Katane;Yousuke Seida;M. Sekine;T. Furuchi;H. Homma
采用RT - PCR技术从秀丽隐杆线虫RNA中分离出4个编码d‐氨基酸氧化酶（DAAO）或d‐天冬氨酸氧化酶（DASPO）的cDNA克隆。克隆的cdna编码的蛋白（Y69Ap、C47Ap、F18Ep和F20Hp）在大肠杆菌中以带N端His标签的重组蛋白表达。除F20Hp外，其余蛋白均从可溶性组分中回收。重组Y69Ap具有功能性DAAO活性，因为它可以脱胺中性和碱性d -氨基酸，而重组C47Ap和F18Ep具有功能性DASPO活性，因为它们可以脱胺酸性d -氨基酸。利用纯化重组蛋白进行的进一步实验表明，Y69Ap对d - Arg的脱氨效率高于d - Ala和d - Met， C47Ap和F18Ep对d - Asp、d - Glu和N -甲基- d - Asp表现出不同的动力学特性。这是首次报道无脊椎动物DAAO和DASPO基因的cDNA克隆。此外，我们的研究首次揭示了秀丽隐杆线虫至少有两个编码功能性DASPOs的基因和一个编码DAAO的基因，尽管以前人们认为生物体只携带这些基因的一个拷贝。两种秀丽隐杆线虫的DASPOs在底物特异性和亚细胞定位上可能存在差异。
Four cDNA clones that were annotated in the database as encoding d‐amino acid oxidase (DAAO) or d‐aspartate oxidase (DASPO) were isolated by RT‐PCR from Caenorhabditis elegans RNA. The proteins (Y69Ap, C47Ap, F18Ep, and F20Hp) encoded by the cloned cDNAs were expressed in Escherichia coli as recombinant proteins with an N‐terminal His‐tag. All proteins except F20Hp were recovered in the soluble fractions. The recombinant Y69Ap has functional DAAO activity, as it can deaminate neutral and basic d‐amino acids, whereas the recombinants C47Ap and F18Ep have functional DASPO activities, as they can deaminate acidic d‐amino acids. Additional experiments using purified recombinant proteins revealed that Y69Ap deaminates d‐Arg more efficiently than d‐Ala and d‐Met, and that C47Ap and F18Ep show distinct kinetic properties against d‐Asp, d‐Glu, and N‐methyl‐d‐Asp. This is the first time that cDNA cloning of invertebrate DAAO and DASPO genes has been reported. In addition, our study reveals for the first time that C. elegans has at least two genes encoding functional DASPOs and one gene encoding DAAO, although it had previously been thought that organisms only bear one copy each of these genes. The two C. elegans DASPOs differ in their substrate specificities and possibly also in their subcellular localization.
登录
查看更多内容
来自牛肾的 D-天冬氨酸氧化酶。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Negri,A;Massey,V;WilliamsJr,CH
通讯作者：
WilliamsJr,CH
D-天冬氨酸对大鼠大细胞神经分泌系统的调节：哺乳动物中天然存在的游离 D-氨基酸的生物学作用的证据。
DOI：
10.1677/joe.0.1670247
发表时间：
2000
期刊：
The Journal of endocrinology
影响因子：
--
作者：
Wang,H;Wolosker,H;Pevsner,J;Snyder,SH;Selkoe,DJ
通讯作者：
Selkoe,DJ
酪氨酸224和酪氨酸228定点突变研究猪肾D-氨基酸氧化酶的动力学机制。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pollegioni,L;Fukui,K;Massey,V
通讯作者：
Massey,V
影响因子：
4.8
作者：
G. D'aniello;A. Tolino;A. D’Aniello;F. Errico;G. Fisher;M. M. Di Fiore-M.
通讯作者：
G. D'aniello;A. Tolino;A. D’Aniello;F. Errico;G. Fisher;M. M. Di Fiore-M.
DOI：
10.1073/pnas.97.9.4926
发表时间：
2000-04-25
影响因子：
11.1
作者：
Mothet, JP;Parent, AT;Snyder, SH
通讯作者：
Snyder, SH