对于β转向显示的最小基架:优化在二硫化物循环的β毛中链的位置
A G Cochran1, R T Tong, M A Starovasnik
1Departments of Protein Engineering and Bioorganic Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, California 94080, USA. andrea@gene.com
Journal of the American Chemical Society
|July 18, 2001
概括
研究人员开发了一种稳定的β-hairpin支架,用于在菌体上显示类库. 这一突破简化了新联体进化的结构-活性关系的定义.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 体显示对于演变的配体至关重要,但理解体构造偏好是具有挑战性的.
- 定义的结构-活性关系是很困难的,因为人们对构造规则的理解很差.
研究的目的:
- 调查二硫化物受约束β-hairpins的结构稳定性.
- 评估β-hairpins作为显示β-turns的支架.
- 简化定义结构-活性关系的过程.
主要方法:
- 用二硫化物形成作为探针测量了19个的折叠的相对自由能量.
- 使用核磁共振 (NMR) 分析了质结构.
- 检查了不同转折序列的酸中的链置换.
主要成果:
- 一种托替代剂显著增强了β-hairpin折叠稳定性.
- 核磁共振数据证实了测量的能量与β-hairpin结构之间的强烈相关性.
- 确定了线性自由能量关系,表明旋转和链-链相互作用对稳定的独立贡献.
- 使用天然的L-氨基酸开发了一个高度稳定的,小的β-hairpin支架.
结论:
- 一个结构化的β转向支架已成功开发用于菌体显示.
- 这种支架表现出有限的构造多样性,适合显示类库.
- 这些发现简化了在进化中定义结构-活性关系的定义.
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