通过 SAM 基质成熟酶的多种乙烯宏环化
Karsten A S Eastman1,2, Andrew G Roberts1, Vahe Bandarian1,2
1Department of Chemistry, The University of Utah, Salt Lake City, UT 84112.
概括
在中,包括含有非天然氨基酸的氨基酸中,PapB酶会产生稳定的乙烯宏循环. 这扩大了用于治疗和化学生物学应用的受约束的设计选择.
科学领域:
- 生物化学
- 化学生物学
- 生物技术
背景情况:
- 二硫化键稳定,但在生理条件下不稳定,限制了生物技术应用.
- PapB是一种S- 腺- L- 甲酶,在成熟过程中通常形成链.
- 通过核糖体产生和翻译后修改的需要稳定策略.
研究的目的:
- 调查PapB的基质范围和酶活性超出其已知的功能.
- 探索PapB在使用各种氨基酸基质形成新型乙烯宏循环中的潜力.
- 扩大酶工具箱以创建受结构约束的.
主要方法:
- 使用PapB与包括D和β氨基酸在内的各种基质进行酶分析.
- 对产物进行分析,以确认α-和β-thioether宏循环的形成.
- 对基体内的非自然氨基酸的PapB耐受性的表征.
主要成果:
- PapB有效地将内部Cys硫醇与以D或β-氨基酸结尾的酸碳酸盐结合在一起,形成α或β-乙烯宏循环.
- PapB对β-和N-甲基氨基酸具有耐受性,使其能够合成完全由非天然氨基酸组成的宏循环.
- 在C端宏循环中,PapB作为序列无关的乙烯连接酶起作用.
结论:
- PapB是一种多功能酶,能够与多种基质形成稳定的乙烯宏循环,包括非天然的氨基酸.
- 这些发现确立了PapB作为构造受结构约束的有价值的工具.
- 这项研究扩大了基于的疗法和化学生物学的可能性.
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