Protein-peptide interactions as probed by tryptophan fluorescence: serum albumins and enkephalin metabolites.

Binding of Leu-enkephalin and the enkephalin metabolite, tyrosine-glycine-glycine (TGG), to bovine serum albumin (BSA) was studied as a model to investigate protein peptide interactions. TGG and Leu-enkephalin quench the tryptophyl fluorescence of BSA. Stern-Volmer quenching constants were typically in the range of 40 to 300 M-1, depending on the experimental conditions. The addition of Cu(II) or Ni(II) did not change the quenching constant, indicating that TGG does not compete for the metal binding sites on BSA. From fluorescence quenching studies with TGG, tyrosyl-glycine, tyrosine and glycyl-glycine, it was concluded that the presence of the tyrosine residue is required for the observed quenching. The phenolic group in tyrosine accounted for the quenching mechanism because phenol was efficient in quenching BSA fluorescence, whereas phenylalanine had no detectable effect. A large solvent isotope effect on the quenching constant of phenol and TGG with BSA strongly suggests an active role of the -OH functionality in the quenching mechanism.