IMIS | Vlaams Instituut voor de Zee
 

Vlaams Instituut voor de Zee

Platform voor marien onderzoek

IMIS

Publicaties | Instituten | Personen | Datasets | Projecten | Kaarten
[ meld een fout in dit record ]mandje (0): toevoegen | toon Print deze pagina

Probing dimerization and intraprotein fluorescence resonance energy transfer in a far-red fluorescent protein from the sea anemone Heteractis crispa
Lessard, G.A.; Habuchi, S.; Werner, J.H.; Goodwin, P.M.; De Schryver, F.; Hofkens, J.; Cotlet, M. (2008). Probing dimerization and intraprotein fluorescence resonance energy transfer in a far-red fluorescent protein from the sea anemone Heteractis crispa. Journal of Biomedical Optics 13(3). dx.doi.org/10.1117/1.2937477
In: Journal of Biomedical Optics. SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS: Bellingham. ISSN 1083-3668; e-ISSN 1560-2281, meer
Peer reviewed article  

Beschikbaar in  Auteurs 

Trefwoord
    Marien
Author keywords
    green fluorescent proteins; HcRed; dimer; fluorescence resonance energytransfer; fluorescence microscopy; single-molecule methods

Auteurs  Top 
  • Lessard, G.A.
  • Habuchi, S.
  • Werner, J.H.
  • Goodwin, P.M.
  • De Schryver, F.
  • Hofkens, J.
  • Cotlet, M.

Abstract
    Proteins from Anthozoa species are homologous to the green fluorescent protein (GFP) from Aequorea victoria but with absorption/emission properties extended to longer wavelengths. HcRed is a far-red fluorescent protein originating from the sea anemone Heteractis crispa with absorption and emission maxima at 590 and 650 nm respectively. We use ultrasensitive fluorescence spectroscopic methods to demonstrate that HcRed occurs as a dimer in solution and to explore the interaction between chromophores within such a dimer. We show that red chromophores within a dimer interact through a Förster-type fluorescence resonance energy transfer (FRET) mechanism. We present spectroscopic evidence for the presence of a yellow chromophore, an immature form of HcRed. This yellow chromophore is involved in directional FRET with the red chromophore when both types of chromophores are part of one dimer. We show that by combining ensemble and single molecule methods in the investigation of HcRed, we are able to sort out subpopulations of chromophores with different photophysical properties and to understand the mechanism of interaction between such chromophores. This study will help in future quantitative microscopy investigations that use HcRed as a fluorescent marker.

Alle informatie in het Integrated Marine Information System (IMIS) valt onder het VLIZ Privacy beleid Top | Auteurs