Department of Medical Biotechnology, College of Biomedical Science, Kangwon National University, Chuncheon, Gangwon-do, 200-701, South Korea.
Protein J. 2013 Jan;32(1):39-47. doi: 10.1007/s10930-012-9457-y.
In the present study, we isolated a methionine sulfoxide reductase B gene, termed SlMSRB1, from tomato (Solanum lycopersicum). In the organ-specific analysis, high expression levels of SlMSRB1 were detected in red mature fruits, leaves and flowers while low transcriptional levels of SlMSRB1 mRNA were observed in stems and roots. In the green fluorescence analysis of SlMSRB1- overexpressed Arabidopsis, signal corresponding to SlMSRB1 was merely detected in chloroplast, suggesting that tomato MSRB1 is a chloroplastial localization protein. Substrate specificity analysis of recombinant SlMSRB1 showed that the enzyme was only targeted to the R epimer of methionine sulfoxide (MetSO) and was able to convert both free and protein-bound MetSO back to methionine in the presence of dithithreitol (DTT). In addition, SlMSRB1 exhibited no activity in thioredoxin dependent system or the substitution of cysteine at position 181 in the DTT-dependent reduction system. Finally, overexpression of SlMSRB1 in yeast revealed that the SlMSRB1 gene might play a critical role in protecting Saccharomyces cerevisiae against oxidative stress.
在本研究中,我们从番茄(Solanum lycopersicum)中分离出一种蛋氨酸亚砜还原酶 B 基因,命名为 SlMSRB1。在组织特异性分析中,SlMSRB1 在红色成熟果实、叶片和花朵中表达水平较高,而在茎和根中 SlMSRB1mRNA 的转录水平较低。在 SlMSRB1 过表达拟南芥的绿色荧光分析中,仅在叶绿体中检测到与 SlMSRB1 对应的信号,表明番茄 MSRB1 是一种叶绿体定位蛋白。重组 SlMSRB1 的底物特异性分析表明,该酶仅靶向蛋氨酸亚砜(MetSO)的 R 对映异构体,并且在存在二硫苏糖醇(DTT)的情况下能够将游离和结合于蛋白质的 MetSO 还原回蛋氨酸。此外,SlMSRB1 在硫氧还蛋白依赖系统或 DTT 依赖还原系统中半胱氨酸 181 位的取代中均没有活性。最后,SlMSRB1 在酵母中的过表达表明 SlMSRB1 基因可能在保护酿酒酵母免受氧化应激方面发挥关键作用。