Protonated form of histidine 238 quenches the fluorescence of tryptophan 241 in subtilisin Novo

Abstract
The pH dependence of the fluorescence of phenylmethanesulfonyl-subtilisin Novo after excitation at 300 nm is studied. pK values of 5.9 and 10.3 were determined from the titration curve. The major source of indole fluorescence is tryptophan-106. Evidence is presented that the protonated form of histidine-238 quenches the fluorescence of tryptophan-241 in this enzyme. The quenching is explained by a charge-transfer complex formation between the excited indole and imidazole rings which are parallel and quite close to each other in the X-ray model.