Photophysical properties of Cerulean and Venus fluorescent proteins

Pabak Sarkar; Srinagesh V. Koushik; Steven S. Vogel; Ignacy Gryczynski; Zygmunt Karol Gryczynski

doi:10.1117/1.3156842

1 May 2009 Photophysical properties of Cerulean and Venus fluorescent proteins

Pabak Sarkar, Srinagesh V. Koushik, Steven S. Vogel, Ignacy Gryczynski, Zygmunt Karol Gryczynski

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Journal of Biomedical Optics, Vol. 14, Issue 3, 034047 (May 2009). https://doi.org/10.1117/1.3156842

Abstract

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.

©(2009) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Pabak Sarkar, Srinagesh V. Koushik, Steven S. Vogel, Ignacy Gryczynski, and Zygmunt Karol Gryczynski "Photophysical properties of Cerulean and Venus fluorescent proteins," Journal of Biomedical Optics 14(3), 034047 (1 May 2009). https://doi.org/10.1117/1.3156842

Published: 1 May 2009

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