Cloning, sequencing, and heterologous expression of rat methionine synthase cDNA.
Cloning, sequencing, and heterologous expression of rat methionine synthase cDNA.
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大鼠蛋氨酸合酶 cDNA 的克隆、测序和异源表达。
DOI：
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
T. Toraya
中科院分区：
文献类型：
--
作者：
K. Yamada;T. Tobimatsu;T. Toraya
蛋氨酸合酶催化钴胺素依赖性甲基转移反应，从 5-甲基四氢叶酸到高半胱氨酸，形成蛋氨酸。通过 RT-PCR、3'-和 5'-RACE 技术克隆并分析了大鼠甲硫氨酸合酶 cDNA。该 cDNA 由 0.3 kb 上游非翻译区、3.8 kb 编码区和 0.4 kb 下游非翻译区组成。该开放阅读框编码1,253个氨基酸残基的多肽，计算分子量为139,162。该分子量与观察到的纯化大鼠肝酶的分子量 (143,000) 非常一致。推导的氨基酸序列分别与大肠杆菌、人类和假定的秀丽隐杆线虫酶的氨基酸序列有 53%、92% 和 64% 的同一性。形成钴胺素和S-腺苷甲硫氨酸结合位点部分的所有指纹序列在大鼠甲硫氨酸合酶中完全保守。通过用含有大鼠甲硫氨酸合酶 cDNA 的杆状病毒感染，在昆虫细胞中实现催化活性酶的高水平表达。
Methionine synthase catalyzes cobalamin-dependent methyl transfer reaction from 5-methyltetrahydrofolate to homocysteine, forming methionine. Rat methonine synthase cDNA was cloned and analyzed by RT-PCR, 3'- and 5'-RACE techniques. The cDNA consists of a 0.3-kb upstream untranslated region, a 3.8-kb coding region, and a 0.4-kb downstream untranslated region. The open reading frame encoded a polypeptide of 1,253 amino acid residues with a calculated molecular weight of 139,162. This molecular weight was in good agreement with the observed one (143,000) of the purified rat liver enzyme. The deduced amino acid sequence was 53, 92, and 64% identical with those of the Escherichia coli, human, and presumptive Caenorhabditis elegans enzymes, respectively. All the fingerprint sequences, forming parts of the cobalamin- and S-adenosylmethionine-binding sites, were completely conserved in the rat methionine synthase. A high-level expression of catalytically active enzyme in insect cells was done by infection with a baculovirus containing the rat methionine synthase cDNA.
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影响因子：
56.9
作者：
DRENNAN, CL;HUANG, S;LUDWIG, ML
通讯作者：
LUDWIG, ML
DOI：
10.1016/s0021-9258(18)80083-7
发表时间：
1989-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Ruma Banerjee;N. L. Johnston;J. K. Sobeski;Prasanta Datta;R. Matthews
通讯作者：
Ruma Banerjee;N. L. Johnston;J. K. Sobeski;Prasanta Datta;R. Matthews
猪肝中哺乳动物蛋氨酸合酶的纯化及其动力学机制。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chen,Z;Crippen,K;Gulati,S;Banerjee,R
通讯作者：
Banerjee,R
影响因子：
3.5
作者：
Li, YN;Gulati, S;Kruger, WD
通讯作者：
Kruger, WD
影响因子：
2.9
作者：
Goulding, CW;Postigo, D;Matthews, RG
通讯作者：
Matthews, RG