双功能的Variediene合成酶的结构为生物合成迪特尔组装和循环利用提供了独特的洞察力
菌双功能烯合成酶具有独特的柱状结构. 研究人员确定了variediene synthase的冷电子显微镜结构,揭示了它的hexameric prenyltransferase核心和循环酶域,提供了对terpenoid生物合成的见解.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 自然产品生物合成 自然产品生物合成
背景情况:
- 菌双功能特尔合成酶是独特的酶,具有连接在一起的prenyltransferase和cyclase域.
- 这些酶通过产生geranylgeranyl diphosphate (GGPP) 和循环它来启动复杂 terpenoids 的生物合成.
- 酶灵活性和弱域相互作用对结构确定构成挑战.
研究的目的:
- 确定大型双功能烯合成酶的第一个实验性衍生结构.
- 为了阐明来自Emericella variecolor*的多种合成酶 (EvVS) 的结构.
- 为了研究双功能烯合成酶中的基质道化.
主要方法:
- 使用冷电子显微镜 (cryo-EM) 来确定495kD EvVS酶的结构.
- 进行了酶学试验,以评估催化域之间的基质通道.
- 基质竞争实验是使用其他酶的循环酶域进行的.
主要成果:
- 冷-EM结构揭示了一个"像杆"的结构,在两个循环酶三合体之间有一个六美式prenyltransferase核心.
- 酵素学数据表明,EvVS.内的prenyltransferase和cyclase域之间缺乏GGPP道.
- 在竞争试验中,来自基甲合成酶的循环酶域优先接受来自EvVS prenyltransferase的GGPP.
结论:
- 虽然EvVS的柱状结构使活跃站点靠近,但这并不能促进内部GGPP的道化.
- EvVS前基转移酶核心的开放侧面表明非本源环酶域结合的潜力,以实现道化.
- 这种结构和机制的洞察力提高了我们对真菌类生物合成和酶工程可能性的理解.
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