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ID: 224661.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
The nature of fluorescence emission in the red fluorescent protein DsRed, revealed by single-molecule detection
Authors:Garcia-Parajo, M. F.; Koopman, M.; van Dijk, E. M. H. P.; Subramaniam, V.; van Hulst, N. F.
Date of Publication (YYYY-MM-DD):2001
Title of Journal:Proceedings of the National Academy of Sciences USA
Journal Abbrev.:Proc. Natl Acad. Sci. U.S.A.
Start Page:14392
End Page:14397
Copyright:Jahrbuch 2002, Copyright MPG 2002
Review Status:Peer-review
Audience:Not Specified
Intended Educational Use:No
Abstract / Description:Recent studies on the newly cloned red fluorescence protein DsRed from the Discosoma genus have shown its tremendous advantages: bright red fluorescence and high resistance against photobleaching. However, it has also become clear that the protein forms closely packed tetramers and there is indication for incomplete protein maturation with unknown proportion of immature green species. We have applied single molecule methodology to elucidate the nature of the fluorescence emission in the DsRed. Real time fluorescence trajectories have been acquired with polarization sensitive detection. Our results indicate that energy transfer between identical monomers occurs efficiently with red emission arising equally likely from any of the chromophoric units. Photodissociation of one of the chromophores weakly quenches the emission of adjacent ones. Dual color excitation (at 488 nm and 568 nm) single molecule microscopy has been performed to reveal the number and distribution of red vs. green species within each tetramer. We find that 86% of the DsRed contain at least one green species with a red-to-green ratio of 1.2 – 1.5. Based on our findings, oligomer suppression would not only be advantageous for protein fusion but it will also increase the fluorescence emission of individual monomers.
Last Change of the Resource (YYYY-MM-DD):2005-08-18
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Applied Optics Group, Department of Applied Physics and MESA Research Institute, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands
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