Ultrafast Electronic and Vibrational Dynamics of Stabilized A State Mutants of the Green Fluorescent Protein (GFP): Snipping the Proton Wire

Deborah Stoner-Ma; Andrew A Jaye; Kate L Ronayne; Jerome Nappa; Peter J Tonge; Stephen R Meech

doi:10.1016/j.chemphys.2008.02.021

Ultrafast Electronic and Vibrational Dynamics of Stabilized A State Mutants of the Green Fluorescent Protein (GFP): Snipping the Proton Wire

Chem Phys. 2008 Jun 23;350(1-3):193-200. doi: 10.1016/j.chemphys.2008.02.021.

Authors

Deborah Stoner-Ma¹, Andrew A Jaye, Kate L Ronayne, Jerome Nappa, Peter J Tonge, Stephen R Meech

Affiliation

¹ Department of Chemistry, Stony Brook University, Stony Brook, New York 11794-3400, USA.

Abstract

Two blue absorbing and emitting mutants (S65G/T203V/E222Q and S65T at pH 5.5) of the green fluorescent protein (GFP) have been investigated through ultrafast time resolved infra-red (TRIR) and fluorescence spectroscopy. In these mutants, in which the excited state proton transfer reaction observed in wild type GFP has been blocked, the photophysics are dominated by the neutral A state. It was found that the A* excited state lifetime is short, indicating that it is relatively less stabilised in the protein matrix than the anionic form. However, the lifetime of the A* state can be increased through modifications to the protein structure. The TRIR spectra show that a large shifts in protein vibrational modes on excitation of the A* state occurs in both these GFP mutants. This is ascribed to a change in H-bonding interactions between the protein matrix and the excited state.

Abstract

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