This paper describes the use of electrospray ionization-Fourier transform ion cyclotron mass spectrometry (ESI-FTICR-MS) to study the relative stabilities of noncovalent complexes of carbonic anhydrase II (CAll, EC 126.96.36.199) and benzenesulfonamide inhibitors in the gas phase. Sustained off-resonance irradiation collision-induced dissociation (SORICID) was used to determine the energetics of dissociation of these CAII-sulfonamide complexes in the gas phase. When two molecules of a benzenesulfonamide (1) were bound simultaneously to one molecule of CAll, one of them was found to exhibit significantly weaker binding (ΔE50 = 0.4 V, where E50 is defined as the amplitude of sustained off-resonance irradiation when 50% of the protein-ligand complexes are dissociated); In Solution, the benzenesulfonamide group coordinates as an anion to a Zn(II)ion bound at the active site of the enzyme. The gas phase stability of the complex with the weakly bound inhibitor was the same as that of the inhibitor complexed with apoCAII (i.e., CAII with the Zn(II) ion removed from the binding site). These results indicate that specific interactions between the sulfonamide group on the inhibitor and the Zn(II)ion on CAII were preserved in the gas phase. Experiments also showed a higher gas phase stability for the complex of para- NO2-benzenesulfonamide-CAII than that for ortho-NO2-benzenesulfonamide- CAII complex. This result further suggests that steric interactions of the inhibitors with the binding pocket of CAll parallel those in Solution. Overall; these results are consistent with the hypothesis that CAll retains, at least partially, the structure of its binding pocket in the gas phase on the time scale (seconds to minutes) of the ESI-FTICR measurements.
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