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Tritium NMR spectroscopy has been used to examine the complexformed by [4-3H]benzenesulfon-amide and human carbonicanhydrase I. The results show that in solution the inhibitor forms a 1:1complex with the enzyme. A 100-spin computational model of the system,constructed with reference to crystallographic results, was used tointerpret tritium relaxation behavior and 3H{1H}NOEs. The analysis shows that the rate of dissociation of theenzyme–sulfonamide complex is 0.35 s−1 and thatthe aromatic ring of the inhibitor undergoes rapid rotation while complexed.