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; Steve 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, Steve Meech, Peter J. Tonge

Research output: Contribution to journal › Article › peer-review

39 Citations (Scopus)

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	English
Pages (from-to)	1321-1328
Number of pages	8
Journal	Biochemistry
Volume	50
Issue number	8
DOIs	https://doi.org/10.1021/bi101589a
Publication status	Published - 10 Jan 2011

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10.1021/bi101589a

Cite this

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., & 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 (AppABLUF) 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 Rui-Kun Zhao and Stelling, {Allison L.} and Richard Brust and Ryu-Ryun Kim 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 Steve Meech and Tonge, {Peter J.}",

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AU - Haigney, Allison

AU - Lukacs, Andras

AU - Zhao, Rui-Kun

AU - Stelling, Allison L.

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, Steve

AU - Tonge, Peter J.

PY - 2011/1/10

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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 (AppABLUF) 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 (AppABLUF) 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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