Proteau Ariane, Shi Rong, Cygler Miroslaw
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Biotechnology Research Institute, Montreal, Quebec, Canada.
Curr Protoc Protein Sci. 2010 Aug;Chapter 17:17.10.1-17.10.9. doi: 10.1002/0471140864.ps1710s61.
Success in determining the three-dimensional structure of a macromolecule by X-ray diffraction methods depends critically on the ability to obtain well ordered crystals of the macromolecule in question. Predisposition to crystallization correlates with the homogeneity of the molecules in solution. Dynamic light scattering (DLS) is particularly well suited for evaluating protein homogeneity under multiple conditions and at concentrations commensurate with crystallization conditions. This unit presents a typical protocol for DLS measurements of a protein sample, and describes approaches to improve protein homogeneity in solution.
通过X射线衍射方法成功测定大分子的三维结构,关键取决于能否获得所研究大分子的有序晶体。结晶倾向与溶液中分子的同质性相关。动态光散射(DLS)特别适合在多种条件下以及与结晶条件相当的浓度下评估蛋白质的同质性。本单元介绍了蛋白质样品DLS测量的典型方案,并描述了提高溶液中蛋白质同质性的方法。