Zhang Tingting, Jin Zhixiong, Zhao Heng, Lai Can, Liu Zheyi, Luo Pan, Dong Zhe, Wang Fangjun
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences Dalian 116023 China
University of Chinese Academy of Sciences Beijing 100049 China.
RSC Adv. 2024 Mar 8;14(12):8075-8080. doi: 10.1039/d3ra08636d. eCollection 2024 Mar 6.
The incorporation of deuterium into peptides and proteins holds broad applications across various fields, such as drug development and structural characterization. Nevertheless, current methods for peptide/protein deuteration often target exchangeable labile sites or require harsh conditions for stable modification. In this study, we present a late-stage approach utilizing an alkaline phosphate solution to achieve deuteration of non-exchangeable backbone sites of peptides and proteins. The specific deuteration regions are identified through ultraviolet photodissociation (UVPD) and mass spectrometry analysis. This deuteration strategy demonstrates site and structure selectivity, with a notable affinity for labeling the α-helix regions of myoglobin. The deuterium method is particularly suitable for peptides and proteins that remain stable under high pH conditions.
将氘掺入肽和蛋白质在药物开发和结构表征等各个领域具有广泛的应用。然而，目前用于肽/蛋白质氘化的方法通常针对可交换的不稳定位点，或者需要苛刻的条件来实现稳定修饰。在本研究中，我们提出了一种后期方法，利用碱性磷酸溶液实现肽和蛋白质不可交换主链位点的氘化。通过紫外光解离（UVPD）和质谱分析确定特定的氘化区域。这种氘化策略表现出位点和结构选择性，对标记肌红蛋白的α-螺旋区域具有显著的亲和力。该氘化方法特别适用于在高pH条件下仍保持稳定的肽和蛋白质。
