| 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，分子量分别为40 kDa和60 kDa。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:Q78 |
| 基金项目:国家自然基金面上项目 |
|
| Screening Proteins Interacting with Sulfide:Quinine Oxidoreductase(SQR) of Urechis unicinctus |
|
tanzhi
|
|
Ocean university of China
|
| Abstract: |
| Sulfide is toxic to most aerobic organisms, even fetal at a level of μmol/L concentration of sulfide in 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, inclusion body of Urechis unicinctus SQR was expressed in E. coli, and obtained activity by protein dilution refolding. Two probable SQR binding proteins, which share molecular weight of 40 ku 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:quinine oxidoreductase His-pulldown mass spectrometer |