在子蛋白胆固醇分析中使用超核基质的通用基质催化物的化学绕道
Daniel A Ciulla1, Michael T Jorgensen2, José-Luis Giner2
1Chemistry Department, Binghamton University (SUNY) , Binghamton, New York 13902, United States.
Journal of the American Chemical Society
|September 21, 2017
概括
鱼蛋白质使用胆固醇进行自我分裂. 一种突变蛋白D303A被用氧化组修饰的胆固醇重新激活,显示出α效应.
科学领域:
- 生物化学
- 分子生物学
- 蛋白质化学
背景情况:
- 刺蛋白质是重要的信号分子.
- 它们通过自我催化内蛋白分解进行自动处理.
- 胆固醇在这个过程中起到核爱的作用,D303被确定为一般的基质催化剂.
研究的目的:
- 调查保存的酸盐残留物 (D303) 在刺自加工中的作用.
- 探索D303激活胆固醇的机制.
- 用替代基质来描述刺突变的活动.
主要方法:
- 在刺蛋白中对D303残留物的局部定向突变.
- 一种新基质3β-氧胆 (3HPC) 的合成.
- 使用胆固醇和3HPC测试突变活性.
主要成果:
- D303转变为氨酸 (D303A) 极大降低了刺自身处理活动 (> 10^4倍).
- D303A突变体与3HPC,一种含有氧基的基质,显著地恢复了活性.
- 其他D303突变 (D303N,D303R,D303E) 也表现出不同程度的3HPC救援活动.
- 3HPC的增强反应性归因于α效应.
结论:
- 刺蛋白中的D303残留物作为胆固醇激活的一般基因催化剂.
- 来自3HPC中的氧化物组的α效应增强了核友性并挽救了突变活性.
- 这项研究提供了对刺自处理机制和基质核友性的理解.
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