Physics
Scientific paper
May 2009
adsabs.harvard.edu/cgi-bin/nph-data_query?bibcode=2009jbo....14c4047s&link_type=abstract
Journal of Journal of Biomedical Optics, Volume 14, Issue 3, pp. 034047-034047-9 (2009).
Physics
Fluorescence, Molecular Biophysics, Molecular Configurations, Proteins, Radiation Quenching
Scientific paper
Cerulean and Venus are recently developed fluorescent proteins, often used as a donor-acceptor pair by researchers in Förster resonance energy transfer-based colocalization studies. We characterized the fluorescent properties of these two proteins in a broad spectral range (form ultraviolet to visible region). Excitation spectra, lifetimes, and polarization spectra show significant energy transfer from aromatic amino acids to the fluorescent protein chromophore. High steady-state anisotropy values and the lack of a fast component in anisotropy decays show that the fluorescent protein chromophore is rigidly fixed within the protein structure. Furthermore, we show that the chromophores are not accessible to external quenchers, such as acrylamide or potassium iodide (KI), allowing the removal of ``unwanted'' background in the environment with external quencher, while leaving the Cerulean/Venus fluorescence unchanged.
Gryczynski Ignacy
Gryczynski Zygmunt
Koushik Srinagesh V.
Sarkar Pabak
Vogel Steven S.
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