Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa

Siarhei Laptenok; Agnieszka A. Gil; Christopher R. Hall; Andras Lukacs; James N. Iuliano; Garth A. Jones; Gregory M. Greetham; Paul Donaldson; Atsushi Miyawaki; Peter J. Tonge; Stephen R. Meech

doi:10.1038/s41557-018-0073-0

Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa

Siarhei Laptenok, Agnieszka A. Gil, Christopher R. Hall, Andras Lukacs, James N. Iuliano, Garth A. Jones, Gregory M. Greetham, Paul Donaldson, Atsushi Miyawaki, Peter J. Tonge, Stephen R. Meech

Biological, Environmental Sciences and Engineering

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

50 Scopus citations

Abstract

Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light-driven structural changes that modulate the fluorescence involve both trans-to-cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are currently not well understood. Here, the mechanism of off-to-on-state switching in dronpa is studied using femtosecond-to-millisecond time-resolved infrared spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore, a ground-state primary product is formed within picoseconds. Surprisingly, the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate, which promotes the final proton-transfer reaction. These data illustrate the interplay between chromophore dynamics and the protein environment underlying fluorescent protein photochromism.

Original language	English (US)
Pages (from-to)	845-852
Number of pages	8
Journal	Nature Chemistry
Volume	10
Issue number	8
DOIs	https://doi.org/10.1038/s41557-018-0073-0
State	Published - Jun 11 2018

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: S.R.M. acknowledges EPSRC for financial support (EP/N033647/1 and EP/M001997/1). P.J.T. acknowledges NSF for financial support (CHE-1223819). A.M. acknowledges the Japan Ministry of Education, Culture, Sports, Science and Technology Grant-in-aid for Scientific research on Innovative Areas: Resonance Bio. The authors acknowledge STFC for access to the Central Laser Facility. Calculations were performed on the High Performance Computing Cluster at the University of East Anglia.

Access to Document

10.1038/s41557-018-0073-0

https://repository.kaust.edu.sa/bitstream/10754/630489/1/Infrared%20sprectroscopy.pdf

Cite this

@article{ea2f7c81fa97462c8def3735a293ba61,

title = "Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa",

abstract = "Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light-driven structural changes that modulate the fluorescence involve both trans-to-cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are currently not well understood. Here, the mechanism of off-to-on-state switching in dronpa is studied using femtosecond-to-millisecond time-resolved infrared spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore, a ground-state primary product is formed within picoseconds. Surprisingly, the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate, which promotes the final proton-transfer reaction. These data illustrate the interplay between chromophore dynamics and the protein environment underlying fluorescent protein photochromism.",

author = "Siarhei Laptenok and Gil, {Agnieszka A.} and Hall, {Christopher R.} and Andras Lukacs and Iuliano, {James N.} and Jones, {Garth A.} and Greetham, {Gregory M.} and Paul Donaldson and Atsushi Miyawaki and Tonge, {Peter J.} and Meech, {Stephen R.}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: S.R.M. acknowledges EPSRC for financial support (EP/N033647/1 and EP/M001997/1). P.J.T. acknowledges NSF for financial support (CHE-1223819). A.M. acknowledges the Japan Ministry of Education, Culture, Sports, Science and Technology Grant-in-aid for Scientific research on Innovative Areas: Resonance Bio. The authors acknowledge STFC for access to the Central Laser Facility. Calculations were performed on the High Performance Computing Cluster at the University of East Anglia.",

year = "2018",

month = jun,

day = "11",

doi = "10.1038/s41557-018-0073-0",

language = "English (US)",

volume = "10",

pages = "845--852",

journal = "Nature Chemistry",

issn = "1755-4330",

publisher = "Nature Publishing Group",

number = "8",

}

TY - JOUR

T1 - Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa

AU - Laptenok, Siarhei

AU - Gil, Agnieszka A.

AU - Hall, Christopher R.

AU - Lukacs, Andras

AU - Iuliano, James N.

AU - Jones, Garth A.

AU - Greetham, Gregory M.

AU - Donaldson, Paul

AU - Miyawaki, Atsushi

AU - Tonge, Peter J.

AU - Meech, Stephen R.

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: S.R.M. acknowledges EPSRC for financial support (EP/N033647/1 and EP/M001997/1). P.J.T. acknowledges NSF for financial support (CHE-1223819). A.M. acknowledges the Japan Ministry of Education, Culture, Sports, Science and Technology Grant-in-aid for Scientific research on Innovative Areas: Resonance Bio. The authors acknowledge STFC for access to the Central Laser Facility. Calculations were performed on the High Performance Computing Cluster at the University of East Anglia.

PY - 2018/6/11

Y1 - 2018/6/11

N2 - Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light-driven structural changes that modulate the fluorescence involve both trans-to-cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are currently not well understood. Here, the mechanism of off-to-on-state switching in dronpa is studied using femtosecond-to-millisecond time-resolved infrared spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore, a ground-state primary product is formed within picoseconds. Surprisingly, the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate, which promotes the final proton-transfer reaction. These data illustrate the interplay between chromophore dynamics and the protein environment underlying fluorescent protein photochromism.

AB - Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light-driven structural changes that modulate the fluorescence involve both trans-to-cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are currently not well understood. Here, the mechanism of off-to-on-state switching in dronpa is studied using femtosecond-to-millisecond time-resolved infrared spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore, a ground-state primary product is formed within picoseconds. Surprisingly, the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate, which promotes the final proton-transfer reaction. These data illustrate the interplay between chromophore dynamics and the protein environment underlying fluorescent protein photochromism.

UR - http://hdl.handle.net/10754/630489

UR - http://link.springer.com/article/10.1038/s41557-018-0073-0

UR - http://www.scopus.com/inward/record.url?scp=85048312421&partnerID=8YFLogxK

U2 - 10.1038/s41557-018-0073-0

DO - 10.1038/s41557-018-0073-0

M3 - Article

SN - 1755-4330

VL - 10

SP - 845

EP - 852

JO - Nature Chemistry

JF - Nature Chemistry

IS - 8

ER -

Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Cite this