TY - JOUR
T1 - Photophysical properties of Cerulean and Venus fluorescent proteins
AU - Sarkar, Pabak
AU - Koushik, Srinagesh V.
AU - Vogel, Steven S.
AU - Gryczynski, Ignacy
AU - Gryczynski, Zygmunt
N1 - Funding Information:
This research was supported by the National Institutes of Health Project No. ZO1AA000452-01 to S.S.V., and by the Texas Emerging Technologies Fund Grant to Z. G.
PY - 2009
Y1 - 2009
N2 - 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.
AB - 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.
KW - Förster resonance energy transfer
KW - fluorescence anisotropy
KW - fluorescence timeline
UR - http://www.scopus.com/inward/record.url?scp=70349263507&partnerID=8YFLogxK
U2 - 10.1117/1.3156842
DO - 10.1117/1.3156842
M3 - Article
C2 - 19566339
AN - SCOPUS:70349263507
SN - 1083-3668
VL - 14
JO - Journal of Biomedical Optics
JF - Journal of Biomedical Optics
IS - 3
M1 - 034047
ER -