Alberta Glycomics Centre and Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada, T6G 2G2.
J Am Soc Mass Spectrom. 2012 Mar;23(3):431-41. doi: 10.1007/s13361-011-0311-9. Epub 2012 Jan 21.
The association-dissociation of noncovalent interactions between protein and ligands, such as other proteins, carbohydrates, lipids, DNA, or small molecules, are critical events in many biological processes. The discovery and characterization of these interactions is essential to a complete understanding of biochemical reactions and pathways and to the design of novel therapeutic agents that may be used to treat a variety of diseases and infections. Over the last 20 y, electrospray ionization mass spectrometry (ESI-MS) has emerged as a versatile tool for the identification and quantification of protein-ligand interactions in vitro. Here, we describe the implementation of the direct ESI-MS assay for the determination of protein-ligand binding stoichiometry and affinity. Additionally, we outline common sources of error encountered with these measurements and various strategies to overcome them. Finally, we comment on some of the outstanding challenges associated with the implementation of the assay and highlight new areas where direct ESI-MS measurements are expected to make significant contributions in the future.
蛋白质与配体（如其他蛋白质、碳水化合物、脂类、DNA 或小分子）之间非共价相互作用的缔合-解离是许多生物过程中的关键事件。发现和描述这些相互作用对于全面了解生化反应和途径以及设计新型治疗剂以治疗各种疾病和感染至关重要。在过去的 20 年中，电喷雾电离质谱（ESI-MS）已成为体外鉴定和定量蛋白质-配体相互作用的多功能工具。在这里，我们描述了直接 ESI-MS 测定法在测定蛋白质-配体结合比和亲和力中的应用。此外，我们还概述了这些测量中常见的误差源以及克服这些误差的各种策略。最后，我们评论了与实施该测定法相关的一些突出挑战，并强调了直接 ESI-MS 测量在未来有望做出重大贡献的新领域。
