Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein

Allison Haigney; Andras Lukacs; Rui Kun Zhao; Allison L. Stelling; Richard Brust; Ryu Ryun Kim; Minako Kondo; Ian Clark; Michael Towrie; Gregory M. Greetham; Boris Illarionov; Adelbert Bacher; Werner Römisch-Margl; Markus Fischer; Stephen R. Meech; Peter J. Tonge

doi:10.1021/bi101589a

Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein

Allison Haigney, Andras Lukacs, Rui Kun Zhao, Allison L. Stelling, Richard Brust, Ryu Ryun Kim, Minako Kondo, Ian Clark, Michael Towrie, Gregory M. Greetham, Boris Illarionov, Adelbert Bacher, Werner Römisch-Margl, Markus Fischer, Stephen R. Meech, Peter J. Tonge

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39 Scopus citations

Abstract

The blue light using flavin (BLUF) domain photosensors, such as the transcriptional antirepressor AppA, utilize a noncovalently bound flavin as the chromophore for photoreception. Since the isoalloxazine ring of the chromophore is unable to undergo large-scale structural change upon light absorption, there is intense interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppA _BLUF) reconstituted with isotopically labeled riboflavin (Rf) and flavin adenine dinucleotide (FAD), which permit the first unambiguous assignment of ground and excited state modes arising directly from the flavin carbonyl groups. Studies of model compounds and DFT calculations of the ground state vibrational spectra reveal the sensitivity of these modes to their environment, indicating that they can be used as probes of structural dynamics.

Original language	British English
Pages (from-to)	1321-1328
Number of pages	8
Journal	Biochemistry
Volume	50
Issue number	8
DOIs	https://doi.org/10.1021/bi101589a
State	Published - 1 Mar 2011

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Haigney, A., Lukacs, A., Zhao, R. K., Stelling, A. L., Brust, R., Kim, R. R., Kondo, M., Clark, I., Towrie, M., Greetham, G. M., Illarionov, B., Bacher, A., Römisch-Margl, W., Fischer, M., Meech, S. R., & Tonge, P. J. (2011). Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein. Biochemistry, 50(8), 1321-1328. https://doi.org/10.1021/bi101589a

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title = "Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein",

abstract = "The blue light using flavin (BLUF) domain photosensors, such as the transcriptional antirepressor AppA, utilize a noncovalently bound flavin as the chromophore for photoreception. Since the isoalloxazine ring of the chromophore is unable to undergo large-scale structural change upon light absorption, there is intense interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppA BLUF) reconstituted with isotopically labeled riboflavin (Rf) and flavin adenine dinucleotide (FAD), which permit the first unambiguous assignment of ground and excited state modes arising directly from the flavin carbonyl groups. Studies of model compounds and DFT calculations of the ground state vibrational spectra reveal the sensitivity of these modes to their environment, indicating that they can be used as probes of structural dynamics.",

author = "Allison Haigney and Andras Lukacs and Zhao, {Rui Kun} and Stelling, {Allison L.} and Richard Brust and Kim, {Ryu Ryun} and Minako Kondo and Ian Clark and Michael Towrie and Greetham, {Gregory M.} and Boris Illarionov and Adelbert Bacher and Werner R{\"o}misch-Margl and Markus Fischer and Meech, {Stephen R.} and Tonge, {Peter J.}",

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T1 - Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein

AU - Haigney, Allison

AU - Lukacs, Andras

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AU - Brust, Richard

AU - Kim, Ryu Ryun

AU - Kondo, Minako

AU - Clark, Ian

AU - Towrie, Michael

AU - Greetham, Gregory M.

AU - Illarionov, Boris

AU - Bacher, Adelbert

AU - Römisch-Margl, Werner

AU - Fischer, Markus

AU - Meech, Stephen R.

AU - Tonge, Peter J.

PY - 2011/3/1

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N2 - The blue light using flavin (BLUF) domain photosensors, such as the transcriptional antirepressor AppA, utilize a noncovalently bound flavin as the chromophore for photoreception. Since the isoalloxazine ring of the chromophore is unable to undergo large-scale structural change upon light absorption, there is intense interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppA BLUF) reconstituted with isotopically labeled riboflavin (Rf) and flavin adenine dinucleotide (FAD), which permit the first unambiguous assignment of ground and excited state modes arising directly from the flavin carbonyl groups. Studies of model compounds and DFT calculations of the ground state vibrational spectra reveal the sensitivity of these modes to their environment, indicating that they can be used as probes of structural dynamics.

AB - The blue light using flavin (BLUF) domain photosensors, such as the transcriptional antirepressor AppA, utilize a noncovalently bound flavin as the chromophore for photoreception. Since the isoalloxazine ring of the chromophore is unable to undergo large-scale structural change upon light absorption, there is intense interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppA BLUF) reconstituted with isotopically labeled riboflavin (Rf) and flavin adenine dinucleotide (FAD), which permit the first unambiguous assignment of ground and excited state modes arising directly from the flavin carbonyl groups. Studies of model compounds and DFT calculations of the ground state vibrational spectra reveal the sensitivity of these modes to their environment, indicating that they can be used as probes of structural dynamics.

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Ultrafast infrared spectroscopy of an isotope-labeled photoactivatable flavoprotein

Abstract

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