Biomedical subjects

M Lei Geng

Publications and source records attributed to M Lei Geng.

3 recordsLinked to original sources

PubMed2006

Generalized two-dimensional heterocorrelation analysis of spectrally resolved and temporally resolved fluorescence of the 8-anilino-1-naphthalenesulfonate-apomyoglobin complex with pH perturbation.

We demonstrate two-dimensional heterocorrelation analysis between spectrally resolved and temporally resolved fluorescence to investigate the decay dynamics of the 8-anilino-1-naphthalenesulfonate- (ANS-) apomyoglobin complex. The dynamic changes of the lifetime components are disclosed across the emission spectrum with an external pH-perturbation. Two different fluorescence lifetime schemes of the ANS-apomyoglobin complex are revealed. From pH 8.5 to 4.5, the transition of protein conformation from the native state to the folding intermediate, a short lifetime component is found to correlate with a short-wavelength emission whose population diminishes with decreasing pH. The lifetime components reflect the excited-state populations of the nascent and the charge-transfer species. From pH 4.2 to 1.0, the transition from the folding intermediate to the acid-unfolded state, the short lifetime is responsible for a long-wavelength emission and the fraction of this component increases when the solution becomes more acidic. In this pH range, the decay components reflect the ground-state populations of microenvironments. The relative decay dynamics across the emission spectrum are revealed without collecting decays at each wavelength. More importantly, these conclusions are reached without the necessity of statistical fitting of the decay data with an a priori decay model.

Algorithms↗

PubMed2006

Analysis of heterogeneous fluorescence decays in proteins. Using fluorescence lifetime of 8-anilino-1-naphthalenesulfonate to probe apomyoglobin unfolding at equilibrium.

The solvatochromic fluorescent dye 8-anilino-1-naphthalenesulfonate (ANS) is one of the popular probes of protein folding. Folding kinetics is tracked with ANS fluorescence intensity, usually interpreted as a reflection of protein structure-the hydrophobicity of the binding environments. Such simplistic view overlooks the complicated nature of ANS-protein complexes: the fluorescence characteristics are convoluted results of the ground state populational distribution of the probe-protein complex, the structural changes in the protein and the excited state photophysics of the probe. Understanding of the interplay of these aspects is crucial in accurate interpretation of the protein dynamics. In this work, the fluorescence decay of ANS complexed with apomyoglobin at different conformations denatured by pH is modeled. The fluorescence decay of the ANS-apomyoglobin complex contains information on not only apomyoglobin structure but also molecular populational distributions. The challenge in modeling fluorescence decay profiles originates from the convolution of heterogeneous binding and excited-state relaxation of the fluorescent probe. We analyzed frequency-domain fluorescence lifetime data of ANS-apomyoglobin with both maximum entropy methods (MEM) and nonlinear least squares methods (NLLS). MEM recovers a model of two expanding-and-merging lifetime distributions for ANS-apomyoglobin in the equilibrium transition from the native (N) through an intermediate (I-1) to the acid-unfolded state U(A). At pH 6.5 and above, when apomyoglobin is mostly populated at the N-state, ANS-apomyoglobin emits a predominant long-lifetime fluorescence from a relaxed charge transfer state S(1,CT) of ANS, and a short-lifetime fluorescence that is mainly from a nascent excited-state S(1,np) of ANS stabilized by the strong ANS-apomyoglobin interaction. Lowering the pH diminishes the contribution from the S(1,np) state. Meanwhile, more protein molecules become populated at the U(A) state, which exhibits a short lifetime that is not distinguishable from the S(1,np) state. At pH 3.4, when the population of the U(A) becomes significant, the short-lifetime fluorescence comes predominantly from ANS binding to the U(A). Further lowering the pH leads to more exposure of the bound ANS. The long lifetime shifts toward and finally merges with the short lifetime and becomes one broad distribution that stands for ANS binding to the U(A) below pH 2.4. The above expanding-and-merging model is consistent with F-statistic analysis of NLLS models. The consistency of this model with the knowledge from the literature, as well as the continuity of the decay parameters changing upon experimental conditions are also crucial in drawing the conclusions.

Anilino Naphthalenesulfonates↗

PubMed2006

Probability-based differential normalized fluorescence bivariate analysis for the classification of tissue autofluorescence spectra.

Differential normalized fluorescence (DNF) is an efficient and effective method for the differentiation of normal and cancerous tissue fluorescence spectra. The diagnostic features are extracted from the difference between the averaged cancerous and averaged normal tissue spectra and used as indices in tissue classification. In this paper, a new method, probability-based DNF bivariate analysis, is introduced based on the univariate DNF method. Two differentiation features are used concurrently in the new method to achieve better classification accuracy. The probability of each sample belonging to a disease state is determined with Bayes decision theory. This probability approach classifies the tissue spectra according to disease states and provides uncertainty information on classification. With a data set of 57 colonic tissue sites, probability-based DNF bivariate analysis is demonstrated to improve the accuracy of cancer diagnosis. The bivariate DNF analysis only requires the collection of a few data points across the entire emission spectrum and has the potential of improving data acquisition speed in tissue imaging.

Analysis of Variance↗