蛋白质组装线组件在神奇酶生物合成:PigA,G,H,I,J的表征
Sylvie Garneau-Tsodikova1, Pieter C Dorrestein, Neil L Kelleher
1Department of Biological Chemistry & Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Journal of the American Chemical Society
|September 28, 2006
概括
研究人员阐明了奇异的生物合成.
科学领域:
- 生物化学 生物化学
- 微生物学 微生物学
- 代谢学 代谢学 代谢学
背景情况:
- 红色代谢物菌体中的一种红色代谢物Prodigiosin,具有独特的三螺旋结构.
- 最初的二烯醇 (A和B环) 形成对于奇异生物合成至关重要.
- 五个关键酶 (PigA,G,H,I,J) 已经涉及到这个二氧化组合.
研究的目的:
- 为了异质地表达和净化参与二氧化形成的五种酶.
- 为了阐明甲醇环A和B生物合成的逐步机制.
- 为了研究PigH酶复合体的翻译后修改.
主要方法:
- 在大肠杆菌中PigA,G,H,I和J的异质表达.
- 五种酶的净化. 五种酶的净化.
- 高性能液体染色学 (HPLC) 和纳米喷雾里埃变换质谱法 (nFTMS) 用于分析.
主要成果:
- 醇环A形成通过两种不同的途径发生,涉及PigI/PigA或Sfp.
- 在PigH转移环A到PigJ后,Pyrrole环B由PigH合成.
- 三个PigH域 (两个ACP和一个SerT) 都经历了翻译后的修改.
结论:
- 详细的机理洞察到2,2-二二烯醇生物合成途径.
- 负责产生神奇素前体的酶机制的表征.
- 了解转化后修改对酶功能的作用.
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