Data from: Native mass spectrometry enabled by infrared matrix-assisted laser desorption electrospray ionization for rapid measurement of protein-ligand biophysical parameters

The determination of equilibrium dissociation constant (Kd) alongside the
maximum binding capacity (Bmax) of protein-ligand interactions is
essential in understanding binding affinities and binding capacities,
which plays a major role in the design and development of new therapeutic
molecules. Rapid and accurate determination of these biophysical
parameters (kd and Bmax) in noncovalent protein – ligand interactions is
desirable in the modern drug discovery process. Previously, we
demonstrated detection of noncovalent protein-ligand complexes by
infrared-assisted matrix desorption electrospray ionization mass
spectrometry (IR-MALDESI-MS). Here, we report the first determination of
biophysical parameters from noncovalent protein - ligand interactions
using a ligand titration approach by IR-MALDESI-MS. Unlike conventional
electrospray ionization-mass spectrometry (ESI-MS), IR-MALDESI-MS enables
analysis in < 13 s per ligand concentration, emphasizing its speed,
automation, and sensitivity. Native mass spectrometry by IR-MALDESI was
performed on carbonic anhydrase II (CAH) incubated with sulfanilamide
(SLFA), which is a known inhibitor. Coupled with other published studies,
these results demonstrate that IR-MALDESI has strong potential as a
high-throughput screening (HTS) technique for rapidly and accurately
determining the Kd and Bmax of protein-ligand complexes.