Hoshino Masaru, Katou Hidenori, Yamaguchi Kei-ichi, Goto Yuji
Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Yoshida-Shimoadachi, Kyoto 606-8501, Japan.
Biochim Biophys Acta. 2007 Aug;1768(8):1886-99. doi: 10.1016/j.bbamem.2007.03.001. Epub 2007 Mar 14.
A general method to analyze the structure of a supramolecular complex of amyloid fibrils at amino acid residue resolution has been developed. This method combines the NMR-detected hydrogen/deuterium (H/D) exchange technique to detect hydrogen-bonded amide groups and the ability of the aprotic organic solvent dimethylsulfoxide (DMSO) to dissolve amyloid fibrils into NMR-observable, monomeric components while suppressing the undesired H/D exchange reaction. Moreover, this method can be generally applied to amyloid fibrils to elucidate the distribution of hydrogen-bonded amino acid residues in the three-dimensional molecular organization in the amyloid fibrils. In this study, we describe theoretical considerations in the H/D exchange method to obtain the structural information of proteins, and the DMSO-quenched H/D exchange method to study a supramolecular complex of amyloid fibrils. A possible application of this method to study the interaction of a protein/peptide with phospholipid membrane is also discussed.
已开发出一种在氨基酸残基分辨率下分析淀粉样纤维超分子复合物结构的通用方法。该方法结合了核磁共振检测的氢/氘（H/D）交换技术来检测氢键结合的酰胺基团，以及非质子有机溶剂二甲基亚砜（DMSO）将淀粉样纤维溶解为核磁共振可观测的单体成分的能力，同时抑制不期望的H/D交换反应。此外，该方法可普遍应用于淀粉样纤维，以阐明淀粉样纤维三维分子结构中氢键结合氨基酸残基的分布。在本研究中，我们描述了用于获取蛋白质结构信息的H/D交换方法以及用于研究淀粉样纤维超分子复合物的DMSO淬灭H/D交换方法的理论考量。还讨论了该方法在研究蛋白质/肽与磷脂膜相互作用方面的可能应用。
