Different phenotypic outcome due to site-specific
phosphorylation in the cancer-associated NQO1 enzyme studied by
phosphomimetic mutations

J 2022

Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations

PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO et. al.

Základní údaje

Originální název

Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations

Autoři

PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO, Petr MAN, Milagros MEDINA, Rogelio PALOMINO-MORALES a Angel L PEY

Vydání

Archives of biochemistry and biophysics, New York, Academic Press, 2022, 0003-9861

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: 3.900

Kód RIV

RIV/00216224:14740/22:00128774

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1016/j.abb.2022.109392

UT WoS

000867178000002

Klíčová slova anglicky

Flavoprotein; Phosphorylation; Structure-function relationships

Štítky

ne MU, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 28. 2. 2023 19:11, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

Protein phosphorylation is a common phenomenon in human flavoproteins although the functional consequences of this site-specific modification are largely unknown. Here, we evaluated the effects of site-specific phosphorylation (using phosphomimetic mutations at sites S40, S82 and T128) on multiple functional aspects as well as in the structural stability of the antioxidant and disease-associated human flavoprotein NQO1 using biophysical and biochemical methods. In vitro biophysical studies revealed effects of phosphorylation at different sites such as decreased binding affinity for FAD and structural stability of its binding site (S82), conformational stability (S40 and S82) and reduced catalytic efficiency and functional cooperativity (T128). Local stability measurements by H/D exchange in different ligation states provided structural insight into these effects. Transfection of eukaryotic cells showed that phosphorylation at sites S40 and S82 may reduce steady-levels of NQO1 protein by enhanced proteasome-induced degradation. We show that site-specific phosphorylation of human NQO1 may cause pleiotropic and counterintuitive effects on this multifunctional protein with potential implications for its relationships with human disease. Our approach allows to establish relationships between site-specific phosphorylation, functional and structural stability effects in vitro and inside cells paving the way for more detailed analyses of phosphorylation at the flavoproteome scale.

Návaznosti

90127, velká výzkumná infrastruktura

Název: CIISB II

Citovat

PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO, Petr MAN, Milagros MEDINA, Rogelio PALOMINO-MORALES a Angel L PEY. Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations. Archives of biochemistry and biophysics. New York: Academic Press, 2022, roč. 729, OCT, s. 109392-109403. ISSN 0003-9861. Dostupné z: https://dx.doi.org/10.1016/j.abb.2022.109392.

@article{2262559,
   author = {PachecoandGarcia, Juan Luis and AnozandCarbonell, Ernesto and Loginov, Dmitry S and Vankova, Pavla and Salido, Eduardo and Man, Petr and Medina, Milagros and PalominoandMorales, Rogelio and Pey, Angel L},
   article_location = {New York},
   article_number = {OCT},
   doi = {http://dx.doi.org/10.1016/j.abb.2022.109392},
   keywords = {Flavoprotein; Phosphorylation; Structure-function relationships},
   language = {eng},
   issn = {0003-9861},
   journal = {Archives of biochemistry and biophysics},
   title = {Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations},
   url = {https://www.sciencedirect.com/science/article/pii/S0003986122002764?via%3Dihub},
   volume = {729},
   year = {2022}
}

TY  - JOUR
ID  - 2262559
AU  - Pacheco-Garcia, Juan Luis - Anoz-Carbonell, Ernesto - Loginov, Dmitry S - Vankova, Pavla - Salido, Eduardo - Man, Petr - Medina, Milagros - Palomino-Morales, Rogelio - Pey, Angel L
PY  - 2022
TI  - Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations
JF  - Archives of biochemistry and biophysics
VL  - 729
IS  - OCT
SP  - 109392
EP  - 109392
PB  - Academic Press
SN  - 00039861
KW  - Flavoprotein
KW  - Phosphorylation
KW  - Structure-function relationships
UR  - https://www.sciencedirect.com/science/article/pii/S0003986122002764?via%3Dihub
N2  - Protein phosphorylation is a common phenomenon in human flavoproteins although the functional consequences of this site-specific modification are largely unknown. Here, we evaluated the effects of site-specific phosphorylation (using phosphomimetic mutations at sites S40, S82 and T128) on multiple functional aspects as well as in the structural stability of the antioxidant and disease-associated human flavoprotein NQO1 using biophysical and biochemical methods. In vitro biophysical studies revealed effects of phosphorylation at different sites such as decreased binding affinity for FAD and structural stability of its binding site (S82), conformational stability (S40 and S82) and reduced catalytic efficiency and functional cooperativity (T128). Local stability measurements by H/D exchange in different ligation states provided structural insight into these effects. Transfection of eukaryotic cells showed that phosphorylation at sites S40 and S82 may reduce steady-levels of NQO1 protein by enhanced proteasome-induced degradation. We show that site-specific phosphorylation of human NQO1 may cause pleiotropic and counterintuitive effects on this multifunctional protein with potential implications for its relationships with human disease. Our approach allows to establish relationships between site-specific phosphorylation, functional and structural stability effects in vitro and inside cells paving the way for more detailed analyses of phosphorylation at the flavoproteome scale.
ER  -

PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO, Petr MAN, Milagros MEDINA, Rogelio PALOMINO-MORALES a Angel L PEY. Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations. \textit{Archives of biochemistry and biophysics}. New York: Academic Press, 2022, roč.~729, OCT, s.~109392-109403. ISSN~0003-9861. Dostupné z: https://dx.doi.org/10.1016/j.abb.2022.109392.

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