Steen Hanno, Jensen Ole Nørregaard
Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
Mass Spectrom Rev. 2002 May-Jun;21(3):163-82. doi: 10.1002/mas.10024.
Photochemical cross-linking is a commonly used method for studying the molecular details of protein-nucleic acid interactions. Photochemical cross-linking aids in defining nucleic acid binding sites of proteins via subsequent identification of cross-linked protein domains and amino acid residues. Mass spectrometry (MS) has emerged as a sensitive and efficient analytical technique for determination of such cross-linking sites in proteins. The present review of the field describes a number of MS-based approaches for the characterization of cross-linked protein-nucleic acid complexes and for sequencing of peptide-nucleic acid heteroconjugates. The combination of photochemical cross-linking and MS provides a fast screening method to gain insights into the overall structure and formation of protein-oligonucleotide complexes. Because the analytical methods are continuously refined and protein structural data are rapidly accumulating in databases, we envision that many protein-nucleic acid assemblies will be initially characterized by combinations of cross-linking methods, MS, and computational molecular modeling.
光化学交联是研究蛋白质 - 核酸相互作用分子细节的常用方法。通过随后鉴定交联的蛋白质结构域和氨基酸残基，光化学交联有助于确定蛋白质的核酸结合位点。质谱（MS）已成为一种灵敏且高效的分析技术，用于确定蛋白质中的此类交联位点。本领域综述描述了许多基于质谱的方法，用于表征交联的蛋白质 - 核酸复合物以及肽 - 核酸杂合物的测序。光化学交联与质谱的结合提供了一种快速筛选方法，以深入了解蛋白质 - 寡核苷酸复合物的整体结构和形成。由于分析方法不断完善且蛋白质结构数据在数据库中迅速积累，我们预计许多蛋白质 - 核酸组装体将首先通过交联方法、质谱和计算分子建模的组合来表征。
