cDNA cloning, expression, and functional characterization of a zebrafish SULT1 cytosolic sulfotransferase
cDNA cloning, expression, and functional characterization of a zebrafish SULT1 cytosolic sulfotransferase
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DOI：
10.1016/s0003-9861(03)00172-3
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发表时间：
2003-06-01
影响因子：
3.9
通讯作者：
Liu, MC
中科院分区：
文献类型：
--
作者：
Sugahara, T;Liu, CC;Liu, MC
利用逆转录-聚合酶链式反应技术，克隆并测定了斑马鱼新的磺基转移酶的全长基因序列。序列分析表明，该基因属于SULT1胞浆硫转移酶基因家族。重组形式的斑马鱼磺基转移酶是从大肠杆菌细胞中纯化出来的，它对许多内源化合物，特别是多巴胺和甲状腺激素，以及包括一些黄酮类、异黄酮类和其他酚类化合物在内的外源化合物显示出硫化活性。斑马鱼磺酸转移酶在最适pH条件下表现出底物依赖性。与以多巴胺为底物测定的结果相比，以没食子酸正丙酯为底物测定的斑马鱼磺基转移酶具有更低的K-m和更高的V-max。热稳定性实验表明，该酶在20至43摄氏度的温度范围内相对稳定。在10种二价金属离子中，Hg2+、Co2+、Zn2+、Cd2+、Cu2+和Pb2+对斑马鱼磺基转移酶的活性有明显的抑制作用。(C)2003年埃尔塞维尔科学公司(美国)。版权所有。
Using the reverse transcriptase-polymerase chain reaction technique, a full-length cDNA encoding a novel zebrafish sulfotransferase was cloned and sequenced. Sequence analysis indicated that this zebrafish sulfotransferase belongs to the SULT1 cytosolic sulfotransferase gene family. The recombinant form of the zebrafish sulfotransferase, purified from Escherichia coli cells, displayed sulfating activities toward a number of endogenous compounds, in particular dopamine and thyroid hormones, in addition to xenobiotics including some flavonoids, isoflavonoids, and other phenolic compounds. The zebrafish sulfotransferase exhibited substrate dependence in pH optimum. In comparison with those determined with dopamine as substrate, the zebrafish sulfotransferase displayed much lower K-m and higher V-max with n-propyl gallate as substrate. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 and 43 degreesC. Among 10 divalent metal cations tested, Hg2+, Co2+, Zn2+, Cd2+, Cu2+, and Pb2+ exhibited dramatic inhibitory effects on the activity of the zebrafish sulfotransferase. (C) 2003 Elsevier Science (USA). All rights reserved.