title: Effect of unfolding on the tryptophanyl fluorescence lifetime distribution in apomyoglobin.
abstract: Proteins exhibit, even in their native state, a large number of conformations differing in small details (substates). The fluorescence lifetime of tryptophanyl residues can reflect the microenvironmental characteristics of these subconformations. We have analyzed the lifetime distribution of the unique indole residue of tuna apomyoglobin (Trp A-12) during the unfolding induced by temperature or guanidine hydrochloride. The results show that the increase of the temperature from 10 to 30 degrees C causes a sharpening of the lifetime distribution. This is mainly due to the higher rate of interconversion among the conformational substates in the native state. A further temperature increase produces partially or fully unfolded states, resulting in a broadening of the tryptophanyl lifetime distribution. The data relative to the guanidine-induced unfolding show a sigmoidal increase of the distribution width, which is due to the transition of the protein structure from the native to the random-coiled state. The broadening of the lifetime distribution indicates that, even in the fully unfolded protein, the lifetime of the tryptophanyl residues is influenced by the protein matrix, which generates very heterogeneous microenvironments.
authors: Cattedra di Chimica e Propedeutica Biochimica, I Facoltà di Medicina e Chirurgia, Università di Napoli, Italy.
authors: E
authors: E
authors: Bismuto
authors: E
authors: E
authors: Gratton
authors: G
authors: G
authors: Irace
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000378:metabolism*; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D007700:Kinetics; D009206:Myocardium / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014364:Tryptophan; D014413:Tuna
chemical_list: D001059:Apoproteins; D006146:Guanidines; D009211:Myoglobin; C010857:apomyoglobin; D014364:Tryptophan; D019791:Guanidine