Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume -, Issue -, Pages -Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jasms.2c00024
Keywords
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Funding
- EPSRC [EP/S002979, EP/R018367/1]
- EPSRC via the Centre for Doctoral Training in Physical Sciences for Health [Sci-Phy-4-Health-EP/L016346/1]
- Alan Turing Institute
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Liquid extraction surface analysis coupled with native mass spectrometry was used to quantitatively analyze protein-ligand interactions. The results showed good consistency when both protein and ligand were present in the sample.
Liquid extraction surface analysis (LESA) coupled to native mass spectrometry (MS) presents unique analytical opportunities due to its sensitivity, speed, and automation. Here, we examine whether this tool can be used to quantitatively probe protein-ligand interactions through calculation of equilibrium dissociation constants (K-d values). We performed native LESA MS analyses for a well-characterized system comprising bovine carbonic anhydrase II and the ligands chlorothiazide, dansylamide, and sulfanilamide, and compared the results with those obtained from direct infusion mass spectrometry and surface plasmon resonance measurements. Two LESA approaches were considered: In one approach, the protein and ligand were premixed in solution before being deposited and dried onto a solid substrate for LESA sampling, and in the second, the protein alone was dried onto the substrate and the ligand was included in the LESA sampling solvent. Good agreement was found between the K-d values derived from direct infusion MS and LESA MS when the protein and ligand were premixed; however, K-d values determined from LESA MS measurements where the ligand was in the sampling solvent were inconsistent. Our results suggest that LESA MS is a suitable tool for quantitative analysis of protein-ligand interactions when the dried sample comprises both protein and ligand.
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