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Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products
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用 (1,2) H 和 (19) F ENDOR 光谱学研究的甲单氧基酶基酶的二铁(III) 和混合价值二铁中心的产品结合.

Stoyan K Smoukov1, Daniel A Kopp, Ann M Valentine

  • 1Department of Chemistry, Northwestern University, Evanston, Illinois 60208, USA.

Journal of the American Chemical Society
|March 14, 2002
PubMed
概括
此摘要是机器生成的。

研究人员使用先进的光谱学研究了乙醇和1,1,1-三乙醇如何与可溶性甲单氧酶 (sMMO) 结合. 他们发现酒精在铁中心附近结合,为酶活性部位相互作用提供了新的见解.

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科学领域:

  • 生物化学 生物化学
  • 酶学 是一种酶学.
  • 频谱学是一种光谱学.

背景情况:

  • 溶性甲单氧酶 (sMMO) 是甲氧化过程中的关键酶.
  • 了解小分子如何与sMMO活性部位相互作用,对于酶机制研究至关重要.
  • sMMO的H ((ox) 和H ((mv) 形式呈现出不同的铁状态,用于研究连接体相互作用.

研究的目的:

  • 研究乙醇和1,1,1-三乙醇 (TFE) 与溶液中的sMMO的H (mv) 和H (ox) 形式的结合.
  • 引入来自TFE的19F核,作为研究小分子与金属酶活性部位结合的新型探针.
  • 为了比较不同smmo氧化还原状态中的酒精的结合方式,并评估dmso的影响.

主要方法:

  • Q频段 (35 GHz) 连续波 (CW) 和脉冲电子核双共振 (ENDOR) 谱学.
  • 使用 (1) H, (2) H 和 (19) F 外源联体 (乙醇,TFE) 的核进行光谱分析.
  • 通过化学还原制备H(mv) 形式,并通过马辐射研究H(ox) 形式,以产生可见于EPR的混合价值二铁中心 ((H(ox)) ((mv)).

主要成果:

  • (19) 结合TFE的F和 (2) H ENDOR光谱,以及结合乙醇的 (1,2) H ENDOR光谱,显示出不同的结合相互作用.
  • 发现酒精与的混合价值中的Fe (II) 离子密切结合.
  • 在H ((ox) 形式中,酒精以桥梁或半桥梁的方式与二铁中心结合.
  • 甲基二硫氧化物 (DMSO) 没有影响乙醇或甲醇与H (ox) 或H (mv) 的结合,但将TFE从H (mv) 的二铁位移.

结论:

  • 这项研究提供了第一个直接证据,证明了酒精对sMMO的溶液状态结合模式.
  • 结果验证了通过将产品扩散到sMMO基酶中获得的晶体结构是溶液结构的代表.
  • 使用 (19) F ENDOR光谱学已被确立为一种有价值的新方法,用于探测金属酶活性位点的配体相互作用.