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大白菜过氧化物还原酶基因新亚型的克隆、表达及其与从同一植株中分离出的2-半胱氨酸过氧化物还原酶的比较

Cloning and expression of a new isotype of the peroxiredoxin gene of Chinese cabbage and its comparison to 2Cys-peroxiredoxin isolated from the same plant.

作者信息

Choi Y O, Cheong N E, Lee K O, Jung B G, Hong C H, Jeong J H, Chi Y H, Kim K, Cho M J, Lee S Y

机构信息

Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Change, 660-701, Korea.

出版信息

Biochem Biophys Res Commun. 1999 May 19;258(3):768-71. doi: 10.1006/bbrc.1999.0714.

Abstract

A cDNA encoding a newly identified isotype of peroxiredoxin (Prx) was isolated from a Chinese cabbage flower bud cDNA library and designated CPrxII. Database searches using the predicted CPrxII amino acid sequence revealed no substantial homology to other proteins with the exception of the yeast type II Prx with which CPrxII shares 27.8% sequence identity. Recombinant CPrxII expressed in Escherichia coli was able to protect glutamine synthetase from inactivation in a metal-catalyzed oxidation system and to reduce H2O2 with electrons provided by thioredoxin. This specific antioxidant activity of CPrxII was about 6-fold higher than that of 2Cys-Prx of the same plant. In contrast to 2Cys-Prx, which is predominantly expressed in leaf tissue of cabbage seedlings, CPrxII is highly expressed in root tissue as revealed by Northern and Western blot analyses. The CPrxII gene exists as a small multigene family in the cabbage genome.

摘要

从大白菜花芽cDNA文库中分离出一个编码新鉴定的过氧化物氧还蛋白(Prx)同型体的cDNA,并将其命名为CPrxII。使用预测的CPrxII氨基酸序列进行数据库搜索发现,除了与CPrxII具有27.8%序列同一性的酵母II型Prx外,与其他蛋白质没有显著同源性。在大肠杆菌中表达的重组CPrxII能够在金属催化的氧化系统中保护谷氨酰胺合成酶不被灭活,并利用硫氧还蛋白提供的电子还原过氧化氢。CPrxII的这种特定抗氧化活性比同一植物的2Cys-Prx高约6倍。与主要在白菜幼苗叶片组织中表达的2Cys-Prx不同,Northern和Western印迹分析表明,CPrxII在根组织中高度表达。CPrxII基因在白菜基因组中以一个小多基因家族的形式存在。

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