On the involvement of electron transfer reactions in the fluorescence decay kinetics heterogeneity of proteins.
Time-resolved fluorescence study of single tryptophan-containing proteins, nuclease, ribonuclease T1, protein G, glucagon, and mastoparan, has been carried out. Three different methods were used for the analysis of fluorescence decays: the iterative reconvolution method, as reviewed and developed in our laboratory, the maximum entropy method, and the recent method that we called "energy transfer" method. All the proteins show heterogeneous fluorescence kinetics (multiexponential decay). The origin of this heterogeneity is interpreted in terms of current theories of electron transfer process, which treat the electron transfer process as a radiationless transition. The theoretical electron transfer rate was calculated assuming the peptide bond carbonyl as the acceptor site. The good agreement between experimental and theoretical electron-transfer rates leads us to suggest that the electron-transfer process is the principal quenching mechanism of Trp fluorescence in proteins, resulting in heterogeneous fluorescence kinetics. Furthermore, the origin of apparent homogeneous fluorescence kinetics (monoexponential decay) in some proteins also can be explained on the basis of electron-transfer mechanism.
Ababou
A
A
Pharmacologie et Physico-Chimie des Interactions Cellulaires et Moléculaires, UMR 7034 CNRS, Université Louis Pasteur, Strasbourg 1, Illkirch, France. axa42@psu.edu
Bombarda
E
E
D001236:Aspergillus oryzae / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry*; D004579:Electron Transport; D005453:Fluorescence; D005934:Glucagon / Q000737:chemistry; D007700:Kinetics; D008836:Micrococcal Nuclease / Q000737:chemistry*; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D006163:Ribonuclease T1 / Q000737:chemistry*; D013211:Staphylococcus aureus / Q000201:enzymology; D013291:Streptococcus / Q000737:chemistry; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry
D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D010455:Peptides; D014364:Tryptophan; D005934:Glucagon; D006163:Ribonuclease T1; D008836:Micrococcal Nuclease