| 摘要: |
| 硫化物是沿海水域中常见的有害物质, 对于多数生物来讲, 其在微摩尔级水平便可导致生物体中毒。硫醌氧化还原酶(SQR)是一种谷胱甘肽还原酶家族的膜结合黄素蛋白, 为硫化物氧化解毒的一种关键酶。本研究以体外原核表达并复性的单环刺螠SQR 为材料, 采用His-pulldown 技术和SDS-PAGE分析, 从单环刺螠体壁细胞裂解液中筛选出两个可能与SQR 结合的蛋白质, 分子质量分别为40 ku 和60 ku。通过毛细管液相色谱-离子肼质谱分析, 初步判断其为细胞色素P450(cytochrome P450)和腺苷三磷酸结合盒转运蛋白(ATP-binding cassette transporter), 并初步探讨了其可能的功能。 |
| 关键词: 单环刺螠 硫醌氧化还原酶 His-pulldown 质谱 |
| DOI： |
| 分类号: |
| 基金项目:国家自然基金面上项目资助（40776074） |
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| Screening proteins interacting with sulfide:quinone oxidoreductase(SQR) of Urechis unicinctus |
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TAN Zhi,MA Yu-bin,SHAO Ming-yu,ZHANG Zhi-feng1
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| Abstract: |
| Sulfide is toxic to most aerobic organisms, even fetal at a level of μmol/L to some organisms. Sulfide: quinone oxidoreductase (SQR), a membrane-bound flavoprotein of glutathione reductase family, is a key enzyme for sulfide oxidation detoxification. In this study, Urechis unicinctus SQR was expressed in Escherichia coli as inclusion bodies, and obtained active proteins by dilution refolding. Two probable SQR-binding proteins, which were of 40 and 60 ku respectively, were screened from the cell lysis solution of U. unicinctus body wall by His-pulldown technology and SDS-PAGE analysis, and preliminary determined as cytochrome P450 (cytochrome P450) and adenosine triphosphate-binding cassette transporter (ATP-binding cassette transporter) by mass spectrometer analysis. Probable functions of the two proteins were discussed. |
| Key words: Urechis unicinctus sulfide:quinone oxidoreductase His-pulldown mass spectrometer |
