Chen Fan, Gülbakan Basri, Weidmann Simon, Fagerer Stephan R, Ibáñez Alfredo J, Zenobi Renato
Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
Institute of Child Health, Division of Pediatric Basic Sciences, Hacettepe University, 06100 Ankara, Turkey.
Mass Spectrom Rev. 2016 Jan-Feb;35(1):48-70. doi: 10.1002/mas.21462. Epub 2015 May 6.
Non-covalent interactions are essential for the structural organization of biomacromolecules and play an important role in molecular recognition processes, such as the interactions between proteins, glycans, lipids, DNA, and RNA. Mass spectrometry (MS) is a powerful tool for studying of non-covalent interactions, due to the low sample consumption, high sensitivity, and label-free nature. Nowadays, native-ESI MS is heavily used in studies of non-covalent interactions and to understand the architecture of biomolecular complexes. However, MALDI-MS is also becoming increasingly useful. It is challenging to detect the intact complex without fragmentation when analyzing non-covalent interactions with MALDI-MS. There are two methodological approaches to do so. In the first approach, different experimental and instrumental parameters are fine-tuned in order to find conditions under which the complex is stable, such as applying non-acidic matrices and collecting first-shot spectra. In the second approach, the interacting species are "artificially" stabilized by chemical crosslinking. Both approaches are capable of studying non-covalently bound biomolecules even in quite challenging systems, such as membrane protein complexes. Herein, we review and compare native-ESI and MALDI MS for the study of non-covalent interactions.
非共价相互作用对于生物大分子的结构组织至关重要，并且在分子识别过程中发挥重要作用，例如蛋白质、聚糖、脂质、DNA和RNA之间的相互作用。质谱（MS）由于样品消耗低、灵敏度高且无需标记，是研究非共价相互作用的强大工具。如今，原生电喷雾电离质谱（native-ESI MS）在非共价相互作用研究以及理解生物分子复合物结构方面被大量使用。然而，基质辅助激光解吸电离质谱（MALDI-MS）也变得越来越有用。当使用MALDI-MS分析非共价相互作用时，检测完整的复合物而不发生碎片化具有挑战性。有两种方法可以做到这一点。在第一种方法中，对不同的实验和仪器参数进行微调，以找到复合物稳定的条件，例如使用非酸性基质并采集首次光谱。在第二种方法中，通过化学交联“人为”稳定相互作用的物种。这两种方法都能够研究非共价结合的生物分子，即使在相当具有挑战性的系统中，如膜蛋白复合物。在此，我们综述并比较原生电喷雾电离质谱和基质辅助激光解吸电离质谱用于非共价相互作用的研究。
