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Distance Measurements Using Partial Donor-Donor Energy Migration within Pairs of Fluorescent Groups in Lipid Bilayers.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
2003 (English)In: Journal of Physical Chemistry B, ISSN 1520-6106, E-ISSN 1520-5207, Vol. 107, no 14, 3318-3324 p.Article in journal (Refereed) Published
Abstract [en]

For distance measurements the fluorescence relaxation was explored within pairwise interacting chemically identical, but photophysically different fluorescent groups. The recently developed theory (Kalinin, S. V.; et al. Spectrochim. Acta, Part A 2002, 58, 1087-1097) of partial donor-donor energy migration (PDDEM) was tested on lipid vesicles as a model system, which, for example, enables arrangement of a pH gradient between the inside and outside of the lipid bilayer. Time-correlated single-photon counting and steady-state fluorescence experiments were performed on mono- and bis-lipid derivatives of fluorescein solubilized in lipid bilayers. The PDDEM approach was successfully applied in the analyses of the different experiments. The distances extracted between the fluorescein groups of bis-fluorescein derivatives in lipid vesicles composed 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-glycero-3-phosphocholine were found to be in reasonable agreement with independent measurements of the bilayer thickness.

Place, publisher, year, edition, pages
2003. Vol. 107, no 14, 3318-3324 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:umu:diva-10055DOI: 10.1021/jp022672cOAI: oai:DiVA.org:umu-10055DiVA: diva2:149726
Available from: 2008-06-12 Created: 2008-06-12 Last updated: 2010-08-05Bibliographically approved
In thesis
1. Electronic Energy Transfer within Asymmetric Pairs of Fluorophores: Partial Donor-Donor Energy Migration (PDDEM)
Open this publication in new window or tab >>Electronic Energy Transfer within Asymmetric Pairs of Fluorophores: Partial Donor-Donor Energy Migration (PDDEM)
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A kinetic model of electronic energy migration within pairs of photophysically non-identical fluorophores has been developed. The model applies to fluorescent groups that exhibit different photophysical and spectral properties when attached to different positions in a macromolecule. The energy migration within such asymmetric pairs is partially reversible, which leads to the case of partial donor-donor energy migration (PDDEM). The model of PDDEM is an extension of the recently developed donor-donor energy migration model (DDEM, F. Bergström et al, PNAS 96 (1999) 12477), and applies to quantitative measurements of energy migration rates and distances within macromolecules. One important distinction from the DDEM model is that the distances can be obtained from fluorescence lifetime measurements. A model of fluorescence depolarisation in the presence of PDDEM is also presented.

To experimentally test the PDDEM approach, different model systems were studied. The model was applied to measure distances between rhodamine and fluorescein groups within on-purpose synthesised molecules that were solubilised in lipid bilayers. Moreover, distances were measured between BODIPY groups in mutant forms of the plasminogen activator inhibitor of type 2 (PAI-2). Measurements of both the fluorescence intensity decays and the time-resolved depolarisation were performed. The obtained distances were in good agreement with independent determinations.

Finally, the PDDEM within pairs of donors is considered, for which both donors exhibit a nonexponential fluorescence decay. In this case it turns out that the fluorescence relaxation of a coupled system contains distance information even if the photophysics of the donors is identical. It is also demonstrated that the choice of relaxation model has a negligible effect on the obtained distances. The latter conclusion holds also for the case of donor-acceptor energy transfer.

Place, publisher, year, edition, pages
Umeå: Kemi, 2004. 36 p.
Keyword
Physical chemistry, fluorescence resonance energy transfer (FRET), donor-donor energy migration (DDEM), homotransfer, fluorescence relaxation, lifetimes, time-resolved fluorescence anisotropy, time-correlated single photon counting, distance measurements, protein structure, Fysikalisk kemi
National Category
Physical Chemistry
Research subject
Physical Chemistry
Identifiers
urn:nbn:se:umu:diva-338 (URN)91-7305-765-7 (ISBN)
Public defence
2004-11-19, KE 32, Kemihuset, Umeå University, Umeå, 13:00 (English)
Opponent
Supervisors
Available from: 2004-10-19 Created: 2004-10-19 Last updated: 2010-08-05Bibliographically approved

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Johansson, Lennart B-Å

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