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工程基板在双功能烯合成酶中道化.

Eliott S Wenger1, Kollin Schultz2,3, Ronen Marmorstein2,4

  • 1Roy and Diana Vagelos Laboratories, Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104-6323.

Proceedings of the National Academy of Sciences of the United States of America
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概括

糖二烯合成酶 (PaFS) 即使没有共价链接器,也能有效地将其 prenyltransferase 和 cyclase 域之间的基质道化. 最佳的道发生在循环酶域与prenyltransferase八合体核心相关联时.

关键词:
冷电子显微镜的使用方法酶催化酶的催化作用蛋白质工程工程 蛋白质工程

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科学领域:

  • 生物化学 生物化学
  • 酶学 是一种酶学.
  • 结构生物学 结构生物学

背景情况:

  • 来自*Phomopsis amygdala*的菌合成酶 (PaFS) 是一种双功能酶.
  • 它拥有前转移酶 (PT) 和循环酶域,由一个灵活的区域连接在一起.
  • PT 域合成了格拉尼尔格拉尼尔二酸盐 (GGPP),而循环酶域将其转化为fusiococcadiene.

研究的目的:

  • 调查链接器和域接近在基质通道中的作用.
  • 确定GGPP在PaFS中的道化结构基础.
  • 评估PaFS PT和循环酶域之间的相互作用偏好.

主要方法:

  • 构建和分析一个"无链接"的PaFS变体.
  • 低温电子显微镜以确定无链接结构的结构.
  • GGPP竞争实验与其他二烯循环类.

主要成果:

  • 对GGPP通道而言,共价联结并不是必不可少的,尽管它可能会提高效率.
  • 无论链接如何,PaFS PT和循环酶域都显示出优先相互作用.
  • 无链接的PaFS结构显示了循环酶域与PT八位体的灵活关联.

结论:

  • 在PaFS中,通过与PT八度体的循环酶域关联来实现最佳的基质通道.
  • 这种关联可以暂时发生或被锁定在位,有或没有共价链接器.
  • 这些发现提供了对多域酶中基质道化机制的见解.