Genetically encoded non-canonical amino acids reveal
asynchronous dark reversion of chromophore, backbone, and
side-chains in EL222

J 2023

Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222

CHAUDHARI, Aditya S; Aditi CHATTERJEE; Catarina A O DOMINGOS; Prokopis C ANDRIKOPOULOS; Yingliang LIU et al.

Základní údaje

Originální název

Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222

Autoři

CHAUDHARI, Aditya S; Aditi CHATTERJEE; Catarina A O DOMINGOS; Prokopis C ANDRIKOPOULOS; Yingliang LIU; Inger ANDERSSON; Bohdan SCHNEIDER; Victor A LORENZ-FONFRIA a Gustavo FUERTES

Vydání

Protein Science, HOBOKEN, WILEY, 2023, 0961-8368

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.500

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:90127/23:00139190

Organizační jednotka

CIISB II

Klíčová slova anglicky

flavoproteins; FTIR spectroscopy; genetic code expansion; kinetics; photosensory receptors; protein structural dynamics; signal transduction; site-specific vibrational probes; time-resolved methods; UV; vis spectroscopy

Štítky

ne MU, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 28. 3. 2025 09:49, Mgr. Eva Dubská

Anotace

V originále

Photoreceptors containing the light-oxygen-voltage (LOV) domain elicit biological responses upon excitation of their flavin mononucleotide (FMN) chromophore by blue light. The mechanism and kinetics of dark-state recovery are not well understood. Here we incorporated the non-canonical amino acid p-cyanophenylalanine (CNF) by genetic code expansion technology at 45 positions of the bacterial transcription factor EL222. Screening of light-induced changes in infrared (IR) absorption frequency, electric field and hydration of the nitrile groups identified residues CNF31 and CNF35 as reporters of monomer/oligomer and caged/decaged equilibria, respectively. Time-resolved multi-probe UV/visible and IR spectroscopy experiments of the lit-to-dark transition revealed four dynamical events. Predominantly, rearrangements around the A'alpha helix interface (CNF31 and CNF35) precede FMN-cysteinyl adduct scission, folding of alpha-helices (amide bands), and relaxation of residue CNF151. This study illustrates the importance of characterizing all parts of a protein and suggests a key role for the N-terminal A'alpha extension of the LOV domain in controlling EL222 photocycle length.

Návaznosti

90127, velká výzkumná infrastruktura

Název: CIISB II

Citovat

CHAUDHARI, Aditya S; Aditi CHATTERJEE; Catarina A O DOMINGOS; Prokopis C ANDRIKOPOULOS; Yingliang LIU; Inger ANDERSSON; Bohdan SCHNEIDER; Victor A LORENZ-FONFRIA a Gustavo FUERTES. Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222. Protein Science. HOBOKEN: WILEY, 2023, roč. 32, č. 4, s. 1-22. ISSN 0961-8368. Dostupné z: https://doi.org/10.1002/pro.4590.

@article{2487199,
   author = {Chaudhari, Aditya S and Chatterjee, Aditi and Domingos, Catarina A O and Andrikopoulos, Prokopis C and Liu, Yingliang and Andersson, Inger and Schneider, Bohdan and LorenzandFonfria, Victor A and Fuertes, Gustavo},
   article_location = {HOBOKEN},
   article_number = {4},
   doi = {https://doi.org/10.1002/pro.4590},
   keywords = {flavoproteins; FTIR spectroscopy; genetic code expansion; kinetics; photosensory receptors; protein structural dynamics; signal transduction; site-specific vibrational probes; time-resolved methods; UV; vis spectroscopy},
   language = {eng},
   issn = {0961-8368},
   journal = {Protein Science},
   title = {Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222},
   url = {https://onlinelibrary.wiley.com/doi/10.1002/pro.4590},
   volume = {32},
   year = {2023}
}

TY  - JOUR
ID  - 2487199
AU  - Chaudhari, Aditya S - Chatterjee, Aditi - Domingos, Catarina A O - Andrikopoulos, Prokopis C - Liu, Yingliang - Andersson, Inger - Schneider, Bohdan - Lorenz-Fonfria, Victor A - Fuertes, Gustavo
PY  - 2023
TI  - Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222
JF  - Protein Science
VL  - 32
IS  - 4
SP  - 1-22
EP  - 1-22
PB  - WILEY
SN  - 09618368
KW  - flavoproteins
KW  - FTIR spectroscopy
KW  - genetic code expansion
KW  - kinetics
KW  - photosensory receptors
KW  - protein structural dynamics
KW  - signal transduction
KW  - site-specific vibrational probes
KW  - time-resolved methods
KW  - UV
KW  - vis spectroscopy
UR  - https://onlinelibrary.wiley.com/doi/10.1002/pro.4590
N2  - Photoreceptors containing the light-oxygen-voltage (LOV) domain elicit biological responses upon excitation of their flavin mononucleotide (FMN) chromophore by blue light. The mechanism and kinetics of dark-state recovery are not well understood. Here we incorporated the non-canonical amino acid p-cyanophenylalanine (CNF) by genetic code expansion technology at 45 positions of the bacterial transcription factor EL222. Screening of light-induced changes in infrared (IR) absorption frequency, electric field and hydration of the nitrile groups identified residues CNF31 and CNF35 as reporters of monomer/oligomer and caged/decaged equilibria, respectively. Time-resolved multi-probe UV/visible and IR spectroscopy experiments of the lit-to-dark transition revealed four dynamical events. Predominantly, rearrangements around the A'alpha helix interface (CNF31 and CNF35) precede FMN-cysteinyl adduct scission, folding of alpha-helices (amide bands), and relaxation of residue CNF151. This study illustrates the importance of characterizing all parts of a protein and suggests a key role for the N-terminal A'alpha extension of the LOV domain in controlling EL222 photocycle length.
ER  -

CHAUDHARI, Aditya S; Aditi CHATTERJEE; Catarina A O DOMINGOS; Prokopis C ANDRIKOPOULOS; Yingliang LIU; Inger ANDERSSON; Bohdan SCHNEIDER; Victor A LORENZ-FONFRIA a Gustavo FUERTES. Genetically encoded non-canonical amino acids reveal asynchronous dark reversion of chromophore, backbone, and side-chains in EL222. \textit{Protein Science}. HOBOKEN: WILEY, 2023, roč.~32, č.~4, s.~1-22. ISSN~0961-8368. Dostupné z: https://doi.org/10.1002/pro.4590.

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