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Analysis of fluorescent spectra from complex biological systems containingvarious fluorescent probes with overlapping emission bands is a challengingtask. Valuable information can be extracted from the full spectra, however, byusing multivariate analysis (MA) of measurements at different wavelengths. Weapplied MA to spectral data of purified Torpedo nicotinic acetylcholinereceptor (AChR) protein reconstituted into liposomes made up ofdioleoylphosphatidic acid (DOPA) and dioleoylphosphatidylcholine (DOPC) dopedwith two extrinsic fluorescent probes (NBD-cholesterol/pyrene-PC). Forsterresonance energy transfer (FRET) was observed between the protein and pyrene-PCand between pyrene-PC and NBD-cholesterol, leading to overlapping emissionbands. Partial least squares analysis was applied to ...

pmc biophysics

Resolution of complex fluorescence spectra of lipids and nicotinic  acetylcholine receptor by multivariate analysis reveals protein-mediated  effects on the receptor's immediate lipid microenvironment