Bourgeois Dominique, Katona Gergely, de Rosny Eve, Carpentier Philippe
Institut de Biologie Structurale Jean-Pierre Ebel, 41 Rue Jules Horowitz, 38027, Grenoble Cedex, France.
Methods Mol Biol. 2009;544:253-67. doi: 10.1007/978-1-59745-483-4_16.
In this chapter, we describe Raman microspectrophotometry applied to crystals of biomolecules. Raman spectra collected in crystallo provide structural information highly complementary to X-ray diffraction, relate the crystalline state to the solution state, and allow the identification of ligand-bound or intermediate states of macromolecules. Nonresonant Raman spectroscopy is particularly suitable to the study of macromolecular crystals, and therefore applies to a wide range of noncolored crystalline proteins. Practical issues related to the investigation of crystals by Raman microspectrophotometry are reviewed, and the current limitations are highlighted.
在本章中,我们描述了应用于生物分子晶体的拉曼显微分光光度法。在晶体中收集的拉曼光谱提供了与X射线衍射高度互补的结构信息,将晶体状态与溶液状态联系起来,并允许识别大分子的配体结合或中间状态。非共振拉曼光谱特别适合于大分子晶体的研究,因此适用于广泛的无色晶体蛋白质。本文综述了通过拉曼显微分光光度法研究晶体的实际问题,并强调了当前的局限性。