Department of Biochemistry, Charles University, Prague, Czech Republic.
Anal Chem. 2013 Feb 5;85(3):1597-604. doi: 10.1021/ac302860m. Epub 2013 Jan 14.
NKR-P1C is an activating immune receptor expressed on the surface of mouse natural killer cells. It has been widely used as a marker for NK cell identification in different mice strains. Recently we solved a crystal structure of the C-type lectin-like domain of a homologous protein, NKR-P1A, using X-ray crystallography and also described the strategy for rapid characterization of the protein conformation in solution. This procedure utilized chemical cross-linking, hydrogen/deuterium exchange, and molecular modeling. It was found that the solution structure differs from the crystal structure in the conformation of the loop region. The loop, detached from the protein compact core in the crystal structure, is closely attached to the core of the protein in solution. Here we present and interpret the solution structure of the C-type lectin-like domain of NKR-P1C using chemical cross-linking and molecular modeling. The validation of the model and conformation of the loop region in NKR-P1C were addressed using ion-mobility mass spectrometry.
NKR-P1C 是一种在小鼠自然杀伤细胞表面表达的激活免疫受体。它已被广泛用作不同小鼠品系中 NK 细胞鉴定的标志物。最近，我们使用 X 射线晶体学解决了同源蛋白 NKR-P1A 的 C 型凝集素样结构域的晶体结构，并描述了用于快速表征蛋白质在溶液中构象的策略。该程序利用化学交联、氢/氘交换和分子建模。结果发现，溶液结构与晶体结构在环区的构象上存在差异。在晶体结构中，从蛋白质紧凑核心上脱离的环区，在溶液中紧密附着在蛋白质核心上。在这里，我们使用化学交联和分子建模来呈现和解释 NKR-P1C 的 C 型凝集素样结构域的溶液结构。使用离子迁移质谱法验证了 NKR-P1C 模型和环区构象的正确性。
