BPTI的支持区域有助于折叠
1Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge 02142.
Cell
|November 27, 1992
概括
牛胰腺素抑制剂 (BPTI) 的pro区域显著加快了蛋白质折叠,增加了产量. 在pro区域中的单个囊蛋白作为分子内试剂,防止错误折叠.
科学领域:
- 生物化学 生物化学
- 蛋白质折叠 蛋白质的折叠
- 分子生物学分子生物学
背景情况:
- 牛胰腺素抑制剂 (BPTI) 折叠在体外是缓慢的,需要几个小时.
- 一半的BPTI分子错误地折叠成非原生中间体.
- 在体内,BPTI与含有氨酸的氨基终端前区域进行合成.
研究的目的:
- 调查BPTI亲区域在蛋白质折叠中的作用.
- 为了确定pro区域的氨酸残留物是否有助于二硫化物键的形成.
- 探索改善BPTI折叠效率的方法.
主要方法:
- 在模型系统中对亲-BPTI的重组表达.
- 在pH7.3,25°C的体外折叠试验.
- 局部定向的突变发生,以评估囊的需求.
- 通过灵活的链接器将单个氨酸连接到BPTI.
主要成果:
- 亲-BPTI显著提高了正确折叠的BPTI的速度和产量在体外.
- 在这种折叠增强过程中,pro区域内的氨酸残留物是必不可少的.
- 一个单一的绑定氨酸足以促进二硫化物键的形成.
- 该pro区域作为一个分子内硫化硫醇试剂.
结论:
- 该BPTI亲区域促进高效和准确的蛋白质折叠.
- 结合的氨酸残留物可以作为分子内试剂来改善二硫化物键的形成.
- 了解这些机制可以为蛋白质工程和治疗性蛋白质生产提供信息.
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