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再组合的Streptomycescoelicolor NiSOD的表达,复制和突变
Peter A Bryngelson1, Sumonu E Arobo, Jennifer L Pinkham
1Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Journal of the American Chemical Society
|January 15, 2004
概括
依赖的超氧化物脱酶 (NiSODs) 提供了一种管理有害超氧化物激素的新方法. 研究表明,改变一个关键的N端胺残留物显著影响NiSOD功能,而不是改变甲胺残留物.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 金属蛋白的研究研究.
背景情况:
- 反应性氧物种 (ROS),特别是超氧化物,会导致细胞损伤.
- 超氧化物脱酶 (SODs) 是一种能中和超氧化物的酶.
- 依赖性SOD (NiSOD) 是这些酶的一个独特类别.
研究的目的:
- 为了表达和描述复合性Streptomyces coelicolor NiSOD的特征.
- 研究特定结联体在NiSOD结构和功能中的作用.
- 阐明NiSOD活动地点的机械和结构方面.
主要方法:
- 这是Streptomyces coelicolor NiSOD的重组表达.
- 酶的体外加工和复制.
- 假定结合残留物 (M28和N-终端H) 的位点定向突变发生.
- 野生类型和突变酶的光谱和催化性能分析.
主要成果:
- 成功生产出完全活跃的重组NiSOD.
- 在位置28 (M28) 的氨酸突变没有影响酶特性.
- 在N端的histidine残留物中发生的突变极大地改变了光谱和催化功能.
- 这些发现凸显了N端的histidine的关键作用.
结论:
- 该N端的histidine对于NiSOD的结构完整性和催化活性至关重要.
- 氨酸28对NiSOD功能没有关键作用.
- 这项研究为NiSODs的新含有活性部位提供了关键的见解.
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