Fluorescence quenching dynamics of tryptophan in proteins. Effect of internal rotation under potential barrier.
In many proteins fluorescence from single tryptophan exhibits a nonexponential decay function. To elucidate the origin of this nonexponential decay, we have examined the fluorescence decay function and time-resolved fluorescence anisotropy of a fluorophore covalently bound to a macromolecule by solving a rotational analogue of the Smoluchowski equation. An angular-dependent quenching constant and potential energy for the fluorophore undergoing internal rotation were introduced into the equation of motion for fluorophore. Results of numerical calculations using the equations thus obtained predict that both the fluorescence decay function and time-resolved anisotropy are dependent on rotational diffusion coefficients of fluorophore and potential energy for the internal rotation. The method was applied to the observed fluorescence decay curve of the single tryptophan in apocytochrome c from horse heart. The calculated decay curves fit the observed ones well.
D000818:Animals; D001059:Apoproteins; D003574:Cytochrome c Group; D045304:Cytochromes c; D003576:Electron Transport Complex IV; D005454:Fluorescence Polarization / Q000379:methods; D006736:Horses; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D009206:Myocardium / Q000201:enzymology; D011506:Proteins; D014364:Tryptophan
D001059:Apoproteins; D003574:Cytochrome c Group; D011506:Proteins; D014364:Tryptophan; D045304:Cytochromes c; D003576:Electron Transport Complex IV