Cloning, expression and analysis of the olfactory glutathione S-transferases in coho salmon.
Cloning, expression and analysis of the olfactory glutathione S-transferases in coho salmon.
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DOI：
10.1016/j.bcp.2012.11.018
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发表时间：
2013-03-15
影响因子：
5.8
通讯作者：
Gallagher, Evan P.
中科院分区：
文献类型：
--
作者：
Espinoza, Herbert M.;Shireman, Laura M.;McClain, Valerie;Atkins, William;Gallagher, Evan P.
谷胱甘肽S-转移酶（GST）通过解毒外源性物质、维持氧化还原状态和调节第二信使来提供细胞保护，所有这些都对维持鲑鱼的嗅觉至关重要。在这里，我们的特点是主要的银鲑嗅觉GST（OlfGST），即欧米茄，PI，和rho子类。OlfGST ω含有一个720 bp的开放阅读框，编码239个氨基酸。OlfGST pi和OlfGST rho分别含有727和681 bp的开放阅读框，编码208和226个氨基酸。全蛋白质质谱分析得出GST omega、pi和rho亚基的分子量分别为29，950、23，354和26，655 Da。同源性建模使用四个蛋白质结构预测算法表明，在所有三个OlfGST异构体的活性位点类似于其他物种的同行。嗅觉GST结合典型的GST底物，但只有OlfGST rho催化农药甲基异硫氰酸酯的脱甲基作用。OlfGST pi和rho表现出对2-羟乙基二硫化物（2-HEDS）和缀合的4-羟基壬烯醛（HNE）（一种具有神经变性性质的毒性醛）的硫醇氧化还原酶活性。OlfGST pi结合HNE的动力学参数为KM = 0.16 ± 0.06 mM和Vmax = 0.5 ± 0.1 μmol min-1 mg-1，而OlfGST rho在催化HNE结合方面更有效（KM = 0.022 ± 0.008 mM和Vmax = 0.47 ± 0.05 μmol min-1 mg-1）。我们的研究结果表明，coho的外周嗅觉系统表达GST异构体，解毒某些亲电试剂和农药，并帮助维持氧化还原状态和信号转导。
The glutathione S-transferases (GSTs) provide cellular protection by detoxifying xenobiotics, maintaining redox status, and modulating secondary messengers, all of which are critical to maintaining olfaction in salmonids. Here, we characterized the major coho salmon olfactory GSTs (OlfGSTs), namely omega, pi, and rho subclasses. OlfGST omega contained an open reading frame of 720 bp and encoded a protein of 239 amino acids. OlfGST pi and OlfGST rho contained open reading frames of 727 and 681 bp, respectively, and encoded proteins of 208 and 226 amino acids. Whole-protein mass spectrometry yielded molecular weights of 29,950, 23,354, and 26,655 Da, respectively, for the GST omega, pi, and rho subunits. Homology modeling using four protein-structure prediction algorithms suggest that the active sites in all three OlfGST isoforms resembled counterparts in other species. The olfactory GSTs conjugated prototypical GST substrates, but only OlfGST rho catalyzed the demethylation of the pesticide methyl parathion. OlfGST pi and rho exhibited thiol oxidoreductase activity towards 2-hydroxyethyl disulfide (2-HEDS) and conjugated 4-hydroxynonenal (HNE), a toxic aldehyde with neurodegenerative properties. The kinetic parameters for OlfGST pi conjugation of HNE were KM = 0.16 ± 0.06 mM and Vmax = 0.5 ± 0.1 μmol min−1 mg−1 for OlfGST pi, whereas OlfGST rho was more efficient at catalyzing HNE conjugation (KM = 0.022 ± 0.008 mM and Vmax = 0.47 ± 0.05 μmol min−1 mg−1). Our findings indicate that the peripheral olfactory system of coho expresses GST isoforms that detoxify certain electrophiles and pesticides and that help maintain redox statusand signal transduction.
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DOI：
10.1016/j.aquatox.2011.12.012
发表时间：
2012-04
期刊：
Aquatic toxicology (Amsterdam, Netherlands)
影响因子：
--
作者：
Espinoza HM;Williams CR;Gallagher EP
通讯作者：
Gallagher EP
影响因子：
3.8
作者：
Gardner, JL;Doi, AM;Gallagher, EP
通讯作者：
Gallagher, EP
影响因子：
2.9
作者：
Balogh, Larissa M.;Le Trong, Isolde;Kripps, Kimberly A.;Shireman, Laura M.;Stenkamp, Ronald E.;Zhang, Wei;Mannervik, Bengt;Atkins, William M.
通讯作者：
Atkins, William M.
影响因子：
3.8
作者：
Abel, EL;Bammler, TK;Eaton, DL
通讯作者：
Eaton, DL
影响因子：
5.8
作者：
Doi, AM;Pham, RT;Gallagher, EP
通讯作者：
Gallagher, EP