umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Electronic Energy Transfer within Asymmetric Pairs of Fluorophores: Partial Donor-Donor Energy Migration (PDDEM)
Umeå University, Faculty of Science and Technology, Department of Chemistry.
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 [en]
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
Keyword [sv]
Fysikalisk kemi
National Category
Physical Chemistry
Research subject
Physical Chemistry
Identifiers
URN: urn:nbn:se:umu:diva-338ISBN: 91-7305-765-7 (print)OAI: oai:DiVA.org:umu-338DiVA: diva2:143149
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
List of papers
1. Partial Donor-Donor Energy Migration (PDDEM) as a Fluorescence Spectroscopic Tool for Measuring Distances in Biomacromolecules
Open this publication in new window or tab >>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, E-ISSN 1873-3557, Vol. 58, no 5, 1087-1097 p.Article in journal (Refereed) Published
Abstract [en]

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

Keyword
Energy migration, Time-resolved fluorescence depolarisation, Intramolecular distances, Biomacromolecules
Identifiers
urn:nbn:se:umu:diva-4159 (URN)doi:10.1016/S1386-1425(01)00613-8 (DOI)
Available from: 2004-10-19 Created: 2004-10-19 Last updated: 2017-12-14Bibliographically approved
2. Distance Measurements Using Partial Donor-Donor Energy Migration within Pairs of Fluorescent Groups in Lipid Bilayers.
Open this publication in new window or tab >>Distance Measurements Using Partial Donor-Donor Energy Migration within Pairs of Fluorescent Groups in Lipid Bilayers.
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.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:umu:diva-10055 (URN)10.1021/jp022672c (DOI)
Available from: 2008-06-12 Created: 2008-06-12 Last updated: 2017-12-14Bibliographically approved
3. Partial donor-donor energy migration (PDDEM): a novel fluorescence method for internal protein distance measurements
Open this publication in new window or tab >>Partial donor-donor energy migration (PDDEM): a novel fluorescence method for internal protein distance measurements
Show others...
2004 (English)In: Physical Chemistry, Chemical Physics - PCCP, ISSN 1463-9076, E-ISSN 1463-9084, Vol. 6, no 11, 3001-3008 p.Article in journal (Refereed) Published
Abstract [en]

We show that the photophysics of chemically identical but photophysically non-identical fluorescent pairs can be used for measuring distances within proteins. For this purpose, the theory of partial donor-donor energy migration (PDDEM, S. Kalinin, J. G. Molotkovsky and L. B.-Angstrom. Johansson, Spectrochim. Acta, Part A, 2002, 58, 1057-1097) was applied for distance measurements between BODIPY groups covalently linked to cystein residues in plasminogen activator inhibitor of type 2 (PAI-2). Two sulfhydryl specific derivatives of BODIPY were used namely: N-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl) iodoacetamide and N-(4.4-difluoro-5.7-ditriethyl-4-bona-3a,4a-diaza-s-indacene-3-yl) methyl iodoacetamide. To determine distances, the time-resolved fluorescence relaxation for two singly labelled forms of PAI-2, as well as the corresponding doubly labelled protein were combined and analysed in a global manner. Fluorescence depolarisation experiments on the labelled mutants were also analysed. The distances determined by PDDEM were in good agreement to those obtained from donor-donor energy migration (DDEM) experiments and structural data on PAI-2. The PDDEM approach allows for the use of very different fluorescent probes, which enables wide range of distances to be measured. The PDDEM model also provides a rational explanation to why previous observations of polyfluorophore-labelled proteins exhibit a shorter average fluorescence lifetime compared to the arithmetic average of lifetimes obtained for the corresponding single labelled proteins.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:umu:diva-14351 (URN)10.1039/b403264k (DOI)
Available from: 2007-06-26 Created: 2007-06-26 Last updated: 2017-12-14Bibliographically approved
4. Energy migration and transfer rates are invariant to modeling the fluorescence relaxation by discrete and continuous distributions of lifetimes
Open this publication in new window or tab >>Energy migration and transfer rates are invariant to modeling the fluorescence relaxation by discrete and continuous distributions of lifetimes
2004 (English)In: Journal of Physical Chemistry B, ISSN 1520-6106, E-ISSN 1520-5207, ISSN 1520-6106, Vol. 108, no 9, 3092-3097 p.Article in journal (Refereed) Published
Abstract [en]

Fluorescent groups typically exhibit nonexponential photophysics when incorporated into biomolecular structures, e.g., proteins and lipid membranes. Models assuming discrete and continuous distributions of lifetimes can each accurately describe the observed relaxation. In the analyses of energy transfer and PDDEM (partial donor-donor energy migration) experiments, one frequently needs to model the nonexponential decays of noninteracting donor and acceptor groups. The present paper aims at exploring whether calculated transfer/migration rates depend on modeling the photophysics' decay by discrete or continuous distributions of lifetimes. Discrete or continuous distribution models of the decay, generated synthetically, were analyzed as well as true experimental data. Two proteins were studied. In one of the systems, we examined energy transfer from Trp (donor) to BODIPY (acceptor) in ribosomal protein S6, obtained from Thermus thermophilus. In the second system, we examined PDDEM between different BODIPY derivatives that were pairwise specifically incorporated in mutant forms of plasminogen activator inhibitor type 2. Interestingly, the rates of electronic energy migration/transfer and distances determined within pairs of interacting chromophores reveal small or negligible influence on using discrete or continuous distributions of lifetimes.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:umu:diva-14319 (URN)10.1021/jp031096x (DOI)
Available from: 2007-06-26 Created: 2007-06-26 Last updated: 2017-12-14Bibliographically approved

Open Access in DiVA

fulltext(834 kB)726 downloads
File information
File name FULLTEXT01.pdfFile size 834 kBChecksum SHA-1
012b104446220220c650bfa2f50f503db56080b0333879a648e3836c430af3cf7967b7dd
Type fulltextMimetype application/pdf

By organisation
Department of Chemistry
Physical Chemistry

Search outside of DiVA

GoogleGoogle Scholar
Total: 726 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 353 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf