PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO, Petr MAN, Milagros MEDINA, Rogelio PALOMINO-MORALES and 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, vol. 729, OCT, p. 109392-109403. ISSN 0003-9861. Available from: https://dx.doi.org/10.1016/j.abb.2022.109392.
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Basic information
Original name Different phenotypic outcome due to site-specific phosphorylation in the cancer-associated NQO1 enzyme studied by phosphomimetic mutations
Authors PACHECO-GARCIA, Juan Luis, Ernesto ANOZ-CARBONELL, Dmitry S LOGINOV, Pavla VANKOVA, Eduardo SALIDO, Petr MAN, Milagros MEDINA, Rogelio PALOMINO-MORALES and Angel L PEY.
Edition Archives of biochemistry and biophysics, New York, Academic Press, 2022, 0003-9861.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10608 Biochemistry and molecular biology
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 3.900
RIV identification code RIV/00216224:14740/22:00128774
Organization unit Central European Institute of Technology
Doi http://dx.doi.org/10.1016/j.abb.2022.109392
UT WoS 000867178000002
Keywords in English Flavoprotein; Phosphorylation; Structure-function relationships
Tags ne MU, rivok
Tags International impact, Reviewed
Changed by Changed by: Mgr. Pavla Foltynová, Ph.D., učo 106624. Changed: 28/2/2023 19:11.
Abstract
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.
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