Okuda Aya, Inoue Rintaro, Morishima Ken, Saio Tomohide, Yunoki Yasuhiro, Yagi-Utsumi Maho, Yagi Hirokazu, Shimizu Masahiro, Sato Nobuhiro, Urade Reiko, Kato Koichi, Sugiyama Masaaki
Institute for Integrated Radiation and Nuclear Science, Kyoto University, Sennan-gun, Osaka 590-0494 Japan.
Institute of Advanced Medical Sciences, Tokushima University, Tokushima 770-8503, Japan.
Biophys Physicobiol. 2021 Feb 6;18:16-27. doi: 10.2142/biophysico.bppb-v18.003. eCollection 2021.
The distinguished feature of neutron as a scattering probe is an isotope effect, especially the large difference in neutron scattering length between hydrogen and deuterium. The difference renders the different visibility between hydrogenated and deuterated proteins. Therefore, the combination of deuterated protein and neutron scattering enables the selective visualization of a target domain in the complex or a target protein in the multi-component system. Despite of this fascinating character, there exist several problems for the general use of this method: difficulty and high cost for protein deuteration, and control and determination of deuteration ratio of the sample. To resolve them, the protocol of protein deuteration techniques is presented in this report. It is strongly expected that this protocol will offer more opportunity for conducting the neutron scattering studies with deuterated proteins.
中子作为散射探针的显著特点是同位素效应,尤其是氢和氘之间中子散射长度的巨大差异。这种差异使得氢化和氘化蛋白质具有不同的可见性。因此,氘化蛋白质与中子散射相结合,能够在复合物中选择性地可视化目标结构域,或在多组分系统中选择性地可视化目标蛋白质。尽管具有这种引人入胜的特性,但该方法的普遍应用仍存在几个问题:蛋白质氘化的难度和高成本,以及样品氘化率的控制和测定。为了解决这些问题,本报告提出了蛋白质氘化技术的方案。强烈期望该方案将为使用氘化蛋白质进行中子散射研究提供更多机会。