Martin Nicholas J, Griffiths Rian L, Edwards Rebecca L, Cooper Helen J
School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
J Am Soc Mass Spectrom. 2015 Aug;26(8):1320-7. doi: 10.1007/s13361-015-1152-8. Epub 2015 May 20.
Liquid extraction surface analysis (LESA) mass spectrometry is a promising tool for the analysis of intact proteins from biological substrates. Here, we demonstrate native LESA mass spectrometry of noncovalent protein complexes of myoglobin and hemoglobin from a range of surfaces. Holomyoglobin, in which apomyoglobin is noncovalently bound to the prosthetic heme group, was observed following LESA mass spectrometry of myoglobin dried onto glass and polyvinylidene fluoride surfaces. Tetrameric hemoglobin [(αβ)2(4H)] was observed following LESA mass spectrometry of hemoglobin dried onto glass and polyvinylidene fluoride (PVDF) surfaces, and from dried blood spots (DBS) on filter paper. Heme-bound dimers and monomers were also observed. The 'contact' LESA approach was particularly suitable for the analysis of hemoglobin tetramers from DBS.
液相萃取表面分析（LESA）质谱法是一种用于分析生物基质中完整蛋白质的很有前景的工具。在此，我们展示了在一系列表面上对肌红蛋白和血红蛋白的非共价蛋白质复合物进行的原位LESA质谱分析。在将肌红蛋白干燥到玻璃和聚偏二氟乙烯表面后进行LESA质谱分析时，观察到了全肌红蛋白，其中脱辅基肌红蛋白与辅基血红素基团非共价结合。在将血红蛋白干燥到玻璃和聚偏二氟乙烯（PVDF）表面以及滤纸上的干血斑（DBS）后进行LESA质谱分析时，观察到了四聚体血红蛋白[（αβ）2（4H）]。还观察到了与血红素结合的二聚体和单体。“接触”LESA方法特别适用于分析来自DBS的血红蛋白四聚体。
