兰西生物合成酶的分布和定向行为通过高分辨率的双重质谱学揭示
M Violet Lee1, Leigh Anne Furgerson Ihnken, Young Ok You
1Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
Journal of the American Chemical Society
|August 12, 2009
概括
兰西生物合成酶LctM和HalM2通过脱水氨基酸和形成兰西环来修改. 哈尔M2主要在N-到C-末端作用,指导着lantibiotic生物合成中的环形成.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 微生物学 微生物学
背景情况:
- 抗菌素是通过核糖体合成的,具有翻译后的修改.
- 兰地生物合成酶是修改前体的关键酶.
- 了解兰西生物合成酶的机制对于新型抗生素的开发至关重要.
研究的目的:
- 阐明了兰地生物合成酶LctM和HalM2.2的酶机制.
- 在体外研究这些酶的脱水和循环活动.
- 为了确定兰因环形成的区域选择性和方向性.
主要方法:
- 里叶变换质谱法 (FTMS) 用于分析酶活动.
- 电子喷雾电离质谱仪 (ESI-MS) 用于观察反应中间体.
- 自由氨酸的化和高分辨率的双重质谱 (MS/MS) 用于区域选择性分析.
主要成果:
- 脱水通过可观测的中间体的分配机制发生.
- 在循环中,HalM2在很大程度上表现出N-至C-终端的方向性.
- 在HalA2基质中由HalM2.2形成了四个氨酸环.
结论:
- 提出了一种兰西生物生产模型,该模型涉及通过N终端领导者结合基质.
- 脱水和循环发生在类似的时间尺度上.
- 该过程显示了N-to-C方向性的高倾向,尽管不是严格绝对的.
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