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Issue 25, 2012
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Excited state proton transfer in strongly enhanced GFP (sGFP2)

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Abstract

Proton transfer is an elementary process in biology. Green fluorescent protein (GFP) has served as an important model system to elucidate the mechanistic details of this reaction, because in GFP proton transfer can be induced by light absorption. We have used pump–dump–probe spectroscopy to study how proton transfer through the ‘proton-wire’ around the chromophore is affected by a combination of mutations in a modern GFP variety (sGFP2). The results indicate that in H2O, after absorption of a photon, a proton is transferred (A* → I*) in 5 ps, and back-transferred from a ground state intermediate (I → A) in 0.3 ns, similar to time constants found with GFPuv, although sGFP2 shows less heterogeneous proton transfer. This suggests that the mutations left the proton-transfer largely unchanged, indicating the robustness of the proton-wire. We used pump–dump–probe spectroscopy in combination with target analysis to probe suitability of the sGFP2 fluorophore for super-resolution microscopy.

Graphical abstract: Excited state proton transfer in strongly enhanced GFP (sGFP2)

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Publication details

The article was received on 05 Mar 2012, accepted on 15 Mar 2012 and first published on 15 Mar 2012


Article type: Paper
DOI: 10.1039/C2CP40694B
Citation: Phys. Chem. Chem. Phys., 2012,14, 8852-8858
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    Excited state proton transfer in strongly enhanced GFP (sGFP2)

    B. van Oort, M. J. T. ter Veer, M. L. Groot and I. H. M. van Stokkum, Phys. Chem. Chem. Phys., 2012, 14, 8852
    DOI: 10.1039/C2CP40694B

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