Photoexcitation of the Blue Light Using FAD Photoreceptor AppA Results in Ultrafast Changes to the Protein Matrix
| Autor: | Peter J. Tonge, Gregory M. Greetham, Allison Haigney, Andras Lukacs, Rui-Kun Zhao, Richard Brust, Michael Towrie, Stephen R. Meech, Ian P. Clark, Allison L. Stelling |
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| Rok vydání: | 2011 |
| Předmět: |
chemistry.chemical_classification
Conformational change Light Molecular Structure Spectrophotometry Infrared Chemistry Infrared Hydrogen bond Carboxylic acid Hydrogen Bonding General Chemistry Flavin group Chromophore Spectrum Analysis Raman Photochemistry Biochemistry Catalysis Photoexcitation Residue (chemistry) Colloid and Surface Chemistry Flavin-Adenine Dinucleotide Photoreceptor Cells |
| Zdroj: | Journal of the American Chemical Society. 133:16893-16900 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/ja2060098 |
| Popis: | Photoexcitation of the flavin chromophore in the BLUF photosensor AppA results in a conformational change that leads to photosensor activation. This conformational change is mediated by a hydrogen-bonding network that surrounds the flavin, and photoexcitation is known to result in changes in the network that include a strengthening of hydrogen bonding to the flavin C4═O carbonyl group. Q63 is a key residue in the hydrogen-bonding network, and replacement of this residue with a glutamate results in a photoinactive mutant. While the ultrafast time-resolved infrared (TRIR) spectrum of Q63E AppA(BLUF) is characterized by flavin carbonyl modes at 1680 and 1650 cm(-1), which are similar in frequency to the analogous modes from the light activated state of the wild-type protein, a band is also observed in the TRIR spectrum at 1724 cm(-1) that is unambiguously assigned to the Q63E carboxylic acid based on U-(13)C labeling of the protein. Light absorption instantaneously (100 fs) bleaches the 1724 cm(-1) band leading to a transient absorption at 1707 cm(-1). Because Q63E is not part of the isoalloxazine electronic transition, the shift in frequency must arise from a sub picosecond perturbation to the flavin binding pocket. The light-induced change in the frequency of the Q63E side chain is assigned to an increase in hydrogen-bond strength of 3 kcal mol(-1) caused by electronic reorganization of the isoalloxazine ring in the excited state, providing direct evidence that the protein matrix of AppA responds instantaneously to changes in the electronic structure of the chromophore and supporting a model for photoactivation of the wild-type protein that involves initial tautomerization of the Q63 side chain. |
| Databáze: | OpenAIRE |
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