Jiang Ruidong, Rempel Don L, Gross Michael L
Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA.
Int J Mass Spectrom. 2023 Aug;490. doi: 10.1016/j.ijms.2023.117080. Epub 2023 May 11.
Although protein footprinting results are commonly obtained by ESI-based LC-MS/MS, a more rapid-turnaround alternative approach is desirable to expand the scope of protein footprinting and facilitate routine analysis such as monitoring protein high order structure in quality control or checking epitope maps. Considering that MALDI is a faster procedure that can be easily adapted for high-throughput analysis, we explore here the feasibility of developing a MALDI-based analysis "portfolio" of bottom-up peptide mass mapping for footprinting. The approach was applied to several model proteins that were submitted to two footprinting strategies, FPOP and GEE labeling, and their performance was evaluated. We found adequate coverage that can be improved with automatic off-line separation and spotting, demonstrating the capability to footprint accurately protein conformational change, showing that MALDI may be useful for selected applications in protein footprinting.
虽然蛋白质足迹分析结果通常通过基于电喷雾电离的液相色谱-串联质谱（ESI-based LC-MS/MS）获得，但为了扩大蛋白质足迹分析的范围并促进常规分析，如在质量控制中监测蛋白质高级结构或检查表位图谱，需要一种周转更快的替代方法。考虑到基质辅助激光解吸电离（MALDI）是一种更快的程序，可轻松适用于高通量分析，我们在此探索开发基于MALDI的自下而上肽质量图谱分析“组合”用于足迹分析的可行性。该方法应用于几种采用两种足迹分析策略（FPOP和GEE标记）的模型蛋白，并评估了它们的性能。我们发现通过自动离线分离和点样可以提高覆盖率，证明了准确测定蛋白质构象变化的能力，表明MALDI可能对蛋白质足迹分析中的特定应用有用。
