Reif Bernd
Munich Center for Integrated Protein Science (CIPSM) at Department Chemie, Technische Universität München, Garching, Germany.
Methods Mol Biol. 2012;831:279-301. doi: 10.1007/978-1-61779-480-3_16.
Perdeuteration and back substitution of exchangeable protons in microcrystalline proteins, in combination with recrystallization from D(2)O-containing buffers, significantly reduce (1)H, (1)H dipolar interactions. This way, amide proton line widths on the order of 20 Hz are obtained. Aliphatic protons are accessible either via specifically protonated precursors or by using low amounts of H(2)O in the bacterial growth medium. The labeling scheme enables characterization of structure and dynamics in the solid-state without dipolar truncation artifacts.
微晶蛋白质中可交换质子的去氘化和回代，结合从含D₂O的缓冲液中重结晶，可显著降低¹H-¹H偶极相互作用。通过这种方式，可获得约20 Hz量级的酰胺质子线宽。脂肪族质子可通过特定质子化前体或在细菌生长培养基中使用少量H₂O来实现标记。该标记方案能够在无偶极截断伪影的情况下对固态中的结构和动力学进行表征。
