Asymmetrical distribution of intrinsic fluorescence lifetimes in proteins

Dynamic fluorescence measurements of proteins in solution are often interpreted in terms of continuous distributions of lifetimes, which reflect the intrinsic structural heterogeneity of these systems. In several cases a single Gaussian or Lorentzian symmetric distribution has been used to fit the data. In this paper we describe a new nonsymmetric Lorentzian function which contains three free parameters (the center, the left, and the right widths) like the double-discrete exponential model (the two lifetimes and one preexponential factor). Simulated data in the frequency domain have been used to compare the fits obtained with these different approaches, introducing a new parameter, ρ, which quantitatively measures the asymmetry of the distribution or the ratio of the two preexponential factors, in the continuous and discrete models, respectively. Real measurements of a mixture of independent fluorophores, as well as of protein fluorescence decays, have also been performed and analyzed in terms of the new asymmetric function. The data have also been fitted with traditional discrete methods (such as the two- and the three-exponential decay) and with another asymmetric function, namely, the skewed Gaussian distribution. © 2002 Plenum Publishing Corporation