Projects per year
Abstract
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 (Kd 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 Kd values derived from direct infusion MS and LESA MS when the protein and ligand were premixed; however, Kd 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.
Original language | English |
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Pages (from-to) | 1168-1175 |
Number of pages | 8 |
Journal | Journal of the American Society for Mass Spectrometry |
Volume | 33 |
Issue number | 7 |
Early online date | 8 Jun 2022 |
DOIs | |
Publication status | Published - 6 Jul 2022 |
Keywords
- Animals
- Carbonic Anhydrase Inhibitors/analysis
- Cattle
- Ligands
- Liquid-Liquid Extraction/methods
- Mass Spectrometry/methods
- Proteins/chemistry
- Solvents
Fingerprint
Dive into the research topics of 'Quantitative Characterization of Three Carbonic Anhydrase Inhibitors by LESA Mass Spectrometry'. Together they form a unique fingerprint.Projects
- 2 Finished
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NAMS: Native ambient mass spectrometry
Engineering & Physical Science Research Council
1/06/19 → 30/11/23
Project: Research
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A new tool to support drug discovery: Native LESA mass spectrometry (NESA)
Engineering & Physical Science Research Council
1/04/18 → 31/03/21
Project: Research Councils
Datasets
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Data to accompany "Quantitative characterisation of three carbonic anhydrase inhibitors by LESA mass spectrometry"
Cooper, H. (Creator), University of Birmingham, 24 Mar 2022
DOI: https://doi.org/10.25500/edata.bham.00000808
Dataset