Findings from University of Maryland Provides New Data about Proteomics (Application of phasor plot and autofluorescence correction for study of heterogeneous cell population)
By a News Reporter-Staff News Editor at Biotech Week -- Data detailed on Proteomics have been presented. According to news originating from Baltimore, Maryland, by NewsRx correspondents, research stated, "Protein-protein interactions in cells are often studied using fluorescence resonance energy transfer (FRET) phenomenon by fluorescence lifetime imaging microscopy (FLIM). Here, we demonstrate approaches to the quantitative analysis of FRET in cell population in a case complicated by a highly heterogeneous donor expression, multiexponential donor lifetime, large contribution of cell autofluorescence, and significant presence of unquenched donor molecules that do not interact with the acceptor due to low affinity of donor-acceptor binding."
Our news journalists obtained a quote from the research from the University of Maryland, "We applied a multifrequency phasor plot to visualize FRET FLIM data, developed a method for lifetime background correction, and performed a detailed time-resolved analysis using a biexponential model. These approaches were applied to study the interaction between the Toll Interleukin-1 receptor (TIR) domain of Toll-like receptor 4 (TLR4) and the decoy peptide 4BB. TLR4 was fused to Cerulean fluorescent protein (Cer) and 4BB peptide was labeled with Bodipy TMRX (BTX). Phasor displays for multifrequency FLIM data are presented. The analytical procedure for lifetime background correction is described and the effect of correction on FLIM data is demonstrated. The absolute FRET efficiency was determined based on the phasor plot display and multifrequency FLIM data analysis."
According to the news editors, the research concluded: "The binding affinity between TLR4-Cer (donor) and decoy peptide 4BB-BTX (acceptor) was estimated in a heterogeneous HeLa cell population."
For more information on this research see: Application of phasor plot and autofluorescence correction for study of heterogeneous cell population. Journal of Biomedical Optics, 2014;19(4):210-218. Journal of Biomedical Optics can be contacted at: Spie-Soc Photo-Optical Instrumentation Engineers, 1000 20TH St, PO Box 10, Bellingham, WA 98225, USA (see also Proteomics).
The news correspondents report that additional information may be obtained from H. Szmacinski, University of Maryland, Sch Med, Dept. of Microbiol & Immunol, Baltimore, MD 21201, United States. Additional authors for this research include V. Toshchakov and J.R. Lakowicz.
Keywords for this news article include: Baltimore, Maryland, United States, North and Central America, Peptides, Proteins, Proteomics
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