Kinetic crystallography by Raman microscopy.
Kinetic crystallography by Raman microscopy.
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DOI：
10.1016/j.bbapap.2010.08.006
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Kalp M
中科院分区：
文献类型：
--
作者：
Carey PR;Chen Y;Gong B;Kalp M
使用拉曼显微镜获得的拉曼光谱，提供了一种独特而敏锐的方法来跟踪浸泡或浸泡条件下单晶内部的相互作用和反应。这种方法的实用性源于发现单个大分子晶体的拉曼光谱在正常（非共振）条件下非常稳定，具有低“光背景”，并为拉曼差分光谱提供了理想的平台。反过来，这使得在非常大而复杂的大分子环境中对亚分子变化的研究成为可能。通常与x射线晶体学有很大的协同作用，拉曼光谱仪为晶体学同事提供最佳的浸泡条件，以产生用于闪冷冻和x射线分析的目标中间体。另一方面，沿着反应路径的点的x射线结构为解释拉曼数据提供了宝贵的基准，这些数据来自拉曼观测到的实时变化的种群。这些原理将通过两个反应来说明：第一个涉及一个复杂的分支反应途径，其基础是临床重要药物化合物对β-内酰胺酶的抑制，其中药物和酶的不同组合在途径的不同区域起作用。第二张图显示了如何从RNA合成起始步骤的几个事件中获得时间数据——更具体地说，当一个GTP分子与一个ATP分子结合在115,000道尔顿晶体RNA聚合酶的活性位点形成G•a二聚体时。最后，我们将总结拉曼显微镜在核酸晶体上的扩展以及已经获得的rna酶的信息。
Raman spectra, obtained using a Raman microscope, offer an unique and incisive approach to follow interactions and reactions inside a single crystal under soak-in or soak-out conditions. The utility of this approach derives from the finding that the Raman spectra from single macromolecular crystals, under normal (non-resonance) conditions, are extremely stable, with a low “light background,” and provide ideal platforms for Raman difference spectroscopy. In turn, this allows the interrogation of sub-molecular changes in very large and complex macromolecular environments. There is often great synergy with X-ray crystallography, with the Raman spectroscopist providing crystallography colleagues with the best soak-in conditions to generate a targeted intermediate for flash freezing and X-ray analysis. On the other hand, X-ray structures at points along a reaction pathway provide invaluable benchmarks for interpreting the Raman data from populations seen by Raman to be changing in real-time. These principles will be illustrated by two reactions: The first involves a complex, branching reaction pathway underlying the inhibition of β-lactamases by clinically important pharmaceutical compounds, where different combinations of drug and enzyme function in different regions of the pathway. The second shows how temporal data can be derived for several events in the initiation step of RNA synthesis—more specifically, when one GTP molecule is joined to one ATP molecule to form a G•A dimer in the active site of a 115,000 Dalton crystalline RNA polymerase. Finally, we will summarize the extension of Raman microscopy to nucleic acid crystals and the information that has been obtained for RNA-based enzymes.
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DOI：
10.1073/pnas.86.5.1515
发表时间：
1989-03-01
影响因子：
11.1
作者：
LOPPNOW, GR;BARRY, BA;MATHIES, RA
通讯作者：
MATHIES, RA
影响因子：
5.6
作者：
LORD, RC;YU, NT
通讯作者：
YU, NT
影响因子：
16
作者：
Gleghorn, Michael L.;Davydova, Elena K.;Murakami, Katsuhiko S.
通讯作者：
Murakami, Katsuhiko S.
影响因子：
15
作者：
Adam, Virgile;Carpentier, Philippe;Bourgeois, Dominique
通讯作者：
Bourgeois, Dominique
影响因子：
2.9
作者：
Christian, Eric L.;Anderson, Vernon E.;Harris, Michael E.
通讯作者：
Harris, Michael E.