Soluble expression of biologically active methionine sulfoxide reductase B1 (PaMsrB1) from Carica papaya in Escherichia coli and isolation of its protein targets

文献类型： 外文期刊

第一作者： Shen, Wentao

作者： Shen, Wentao;Han, Jie;Yan, Pu;Zhang, Lie;Li, Xiaoying;Tuo, Decai;Zhou, Peng;Shen, Wentao;Han, Jie;Yan, Pu;Zhang, Lie;Li, Xiaoying;Tuo, Decai;Zhou, Peng;Han, Jie;Zheng, Jiping;Zhang, Lie

作者机构：

关键词： Methionine sulfoxide reductase B1; Chloroplast; Recombinant expression; Interacting protein

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

ISSN： 1046-5928

年卷期： 2018 年 146 卷

页码：

收录情况： SCI

摘要： Plant methionine sulfoxide reductase B1 (MsrB1) protects the photosynthetic apparatus from oxidative damage by scavenging reactive oxygen species to repair Met-oxidized proteins in response to abiotic stresses and biotic attack. Papaya MsrB1 (PaMsrB1) was identified previously to interact with papaya ringspot virus NIa-Pro, and this interaction inhibits the import of PaMsrB1 into the chloroplast. Further functional characterization of PaMsrB1 requires the production of a biologically active purified recombinant protein. In this report, PaMsrB1 as a fusion protein containing an N-terminal maltose-binding protein (MBP) was expressed in Escherichia coil Rosetta (DE3) cells and purified. Production of soluble fusion protein was greater when the cells were cultured at 16 degrees C than at 37 degrees C. The Factor Xa protease digested MBP-PaMsrB1 fusion protein and subsequently purified recombinant PaMsrB1 specifically reduced the R-diastereomer of methionine sulfoxide (MetSO) and Dabsyl-MetSO to Met in the presence of dithiothreitol. Eight chloroplast-localized and five non-chloroplast-localized candidate proteins that interact with PaMsrB1 were isolated by affinity chromatography and liquid chromatography coupled to tandem mass spectrometry. The results provide a platform to further understand the anti oxidative defense mechanism of PaMsrB1.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序