Scavenius Carsten, Ghodke Shirin, Otzen Daniel E, Enghild Jan J
Center for Insoluble Protein Structure and Interdisciplinary Nanoscience Center (iNANO) at the Department of Molecular Biology, Science Park, University of Aarhus, Gustav Wieds Vej 10c, 8000 Aarhus C, Denmark.
Int J Mass Spectrom. 2011 Apr 30;302(1-3):167-173. doi: 10.1016/j.ijms.2010.10.001.
In this study, we have used glucagon as a model system for analyzing amyloid fibrillogenesis by hydrogen exchange MALDI mass spectrometry (HXMS). The hydrogen exchange mass spectrometry data correlated well with the traditional method based on Thioflavin T fluorescence and provided quantitative information by measuring the fibrillating molecules directly. The hydrogen exchange mass spectrometry data collected during fibrillogenesis revealed that glucagon fibrillation was a two component system showing an on/off type of interaction where only monomeric and fibrils were present without any substantial amount of intermediate species. This was evident by the extensive deuteration of the monomer and protection of the entire 29 residue glucagon peptide upon fibrillation.. The method complements the traditional procedures and has the potential to provide new information with respect to the nature of transient species, the structure of the growing fibrils and the mechanism of formation.
在本研究中，我们使用胰高血糖素作为模型系统，通过氢交换基质辅助激光解吸电离质谱法（HXMS）分析淀粉样纤维形成。氢交换质谱数据与基于硫黄素T荧光的传统方法相关性良好，并通过直接测量形成纤维的分子提供了定量信息。在纤维形成过程中收集的氢交换质谱数据表明，胰高血糖素纤维化是一个双组分系统，呈现开/关型相互作用，其中仅存在单体和纤维，没有任何大量的中间物种。这通过单体的广泛氘代和纤维化时整个29个残基的胰高血糖素肽的保护得以证明。该方法补充了传统程序，并且有可能提供关于瞬态物种的性质、生长纤维的结构和形成机制的新信息。
