ATP Changes the Fluorescence Lifetime of Cyan Fluorescent Protein via an Interaction with His148

Petoukhov, Eugenia; DeRose, James

November 05, 2010 Abstract

ATP Changes the Fluorescence Lifetime of Cyan Fluorescent Protein via an Interaction with His148

This study employed fluorescence lifetime spectroscopy to investigate the influence of ATP (adenosine triphosphate) on the fluorescence intensities of yellow and cyan fluorescent proteins (YFP/CFP) which are used as the basis for FRET (Förster resonance energy transfer) sensors. This work further elucidates the complex and heterogeneous fluorescence properties of CFP and its mutant H148D, as well as the sensitivity of CFP fluorescence to changes in solute conditions.

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¹Leica Microsystems

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Previous research has demonstrated that ATP has an effect on YFP/CFP fluorescence intensities observed with CFP-YFP-based FRET sensors. This phenomenon was thought to take place by quenching during the energy transfer step, potentially coupled to energy-induced charge displacement in the phosphate groups.

Using different donor and acceptor pairs, the study found that changes in ATP concentrations affected only the CFP-YFP-based FRET sensors. Using time-resolved fluorescence spectroscopy, the results demonstrate that the effect of ATP is not due to quenching at the energy transfer step, but more likely by a direct interaction of ATP with the positively charged histidine residue at position 148 (His148) of CFP. Gaining a better understanding of the emissive properties of fluorescent proteins might be important for making improved designs of FRET sensors.

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Borst JW, Willemse M, Slijkhuis R, van der Krogt G, Laptenok SP, Jalink K, Wieringa B and Fransen JAM: