Partial Donor-Donor Energy Migration (PDDEM) as a Fluorescence Spectroscopic Tool for Measuring Distances in Biomacromolecules
2002 (English)In: Spectrochimica Acta Part A - Molecular and Biomolecular Spectroscopy, ISSN 1386-1425, Vol. 58, no 5, 1087-1097 p.Article in journal (Refereed) Published
A theoretical model is presented, tested and applied for determining the rates of energy migration and distances within pairs of chemically identical fluorophores, so-called donors (D), which are exposed to different physical properties. The model is a general extension of the recently developed donor–donor energy migration (DDEM) model [J. Chem. Soc., Faraday Trans. 92 (1996)1563; J. Chem. Phys. 105 (1996) 10896] that applies to examining structure-function of biomacromolecules, such as proteins. Most fluorescent groups of the same kind incorporated at different positions (α and β) in a macromolecule exhibit shifts of the absorption and/or emission spectra, as well as different relaxation rates of the photophysics. As a consequence, the energy migration between the Dα and Dβ groups will be partially reversible. We refer to this case, as the partial donor–donor energy migration (PDDEM). The models of PPDEM presented can be used for analysing time-resolved fluorescence relaxation, as well as fluorescence depolarisation experiments. To explore the limitations of the PDDEM model, we have generated and re-analysed synthetic data that mimic time-correlated single photon counting (TCSPC) experiments. It was found that slow and fast rates of energy migration are most accurately recovered from the fluorescence relaxation and the depolarisation experiments, respectively. At comparable transfer and fluorescence rates, both kinds of experiments are equally useful. Real experiments on PDDEM were performed on an asymmetrically quenched bichromophoric molecule (1,32-dihydroxy-dotriacontane-bis-(Rhodamine 101) ester), that spans across the lipid bilayer of a vesicle. The depolarisation data were analysed by the PDDEM model and provide a distance between Rhodamine 101 groups, which agrees with independent studies
Place, publisher, year, edition, pages
2002. Vol. 58, no 5, 1087-1097 p.
Energy migration, Time-resolved fluorescence depolarisation, Intramolecular distances, Biomacromolecules
IdentifiersURN: urn:nbn:se:umu:diva-4159DOI: doi:10.1016/S1386-1425(01)00613-8OAI: oai:DiVA.org:umu-4159DiVA: diva2:143145