Next Article in Journal
Chitosan Nanocomposite Coatings Containing Chemically Resistant ZnO–SnOx Core–shell Nanoparticles for Photocatalytic Antifouling
Previous Article in Journal
Expression Analysis of Zinc Transporters in Nervous Tissue Cells Reveals Neuronal and Synaptic Localization of ZIP4
 
Search for Articles:
Title / Keyword
Author / Affiliation
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
Journals
IJMS
Volume 22
Issue 9
10.3390/ijms22094512
ijms-logo
Submit to this Journal Review for this Journal Edit a Special Issue
► Article Menu

Article Menu

share announcement question_answer thumb_up
...
textsms
...
Article

Effect of Posttranslational Modifications on the Structure and Activity of FTO Demethylase

by
Michał Marcinkowski
1,
Tomaš Pilžys
1,
Damian Garbicz
1,
Jan Piwowarski
1,
Damian Mielecki
1,
Grzegorz Nowaczyk
2,
Michał Taube
3,
Maciej Gielnik
3,
Maciej Kozak
3,4,
Maria Winiewska-Szajewska
1,
Ewa Szołajska
1,
Janusz Dębski
1,
Agnieszka M. Maciejewska
1,
Kaja Przygońska
1,
Karolina Ferenc
5,
Elżbieta Grzesiuk
1,*,† and
Jarosław Poznański
1,*,†
1
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland
2
NanoBioMedical Centre, Adam Mickiewicz University, Wszechnicy Piastowskiej 3, 61-614 Poznan, Poland
3
Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University, Uniwersytetu Poznanskiego 2, 61-614 Poznan, Poland
4
National Synchrotron Radiation Centre SOLARIS, Jagiellonian University, Czerwone Maki 98, 30-392 Kraków, Poland
5
Veterinary Research Centre, Department of Large Animal Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 100, 02-797 Warsaw, Poland
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Academic Editor: Christina Piperi
Int. J. Mol. Sci. 2021, 22(9), 4512; https://doi.org/10.3390/ijms22094512
Received: 12 March 2021 / Revised: 19 April 2021 / Accepted: 22 April 2021 / Published: 26 April 2021
(This article belongs to the Section Molecular Toxicology)
View Full-Text Download PDF
Browse Figures
Review Reports

Abstract

The FTO protein is involved in a wide range of physiological processes, including adipogenesis and osteogenesis. This two-domain protein belongs to the AlkB family of 2-oxoglutarate (2-OG)- and Fe(II)-dependent dioxygenases, displaying N6-methyladenosine (N6-meA) demethylase activity. The aim of the study was to characterize the relationships between the structure and activity of FTO. The effect of cofactors (Fe2+/Mn2+ and 2-OG), Ca2+ that do not bind at the catalytic site, and protein concentration on FTO properties expressed in either E. coli (ECFTO) or baculovirus (BESFTO) system were determined using biophysical methods (DSF, MST, SAXS) and biochemical techniques (size-exclusion chromatography, enzymatic assay). We found that BESFTO carries three phosphoserines (S184, S256, S260), while there were no such modifications in ECFTO. The S256D mutation mimicking the S256 phosphorylation moderately decreased FTO catalytic activity. In the presence of Ca2+, a slight stabilization of the FTO structure was observed, accompanied by a decrease in catalytic activity. Size exclusion chromatography and MST data confirmed the ability of FTO from both expression systems to form homodimers. The MST-determined dissociation constant of the FTO homodimer was consistent with their in vivo formation in human cells. Finally, a low-resolution structure of the FTO homodimer was built based on SAXS data. View Full-Text
Keywords: ECFTO; BESFTO; phosphorylation; calcium; dimerization; nanoDSF; MST; HDX; SAXS ECFTO; BESFTO; phosphorylation; calcium; dimerization; nanoDSF; MST; HDX; SAXS
►▼ Show Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

Supplementary Material

SciFeed

Share and Cite

MDPI and ACS Style

Marcinkowski, M.; Pilžys, T.; Garbicz, D.; Piwowarski, J.; Mielecki, D.; Nowaczyk, G.; Taube, M.; Gielnik, M.; Kozak, M.; Winiewska-Szajewska, M.; Szołajska, E.; Dębski, J.; Maciejewska, A.M.; Przygońska, K.; Ferenc, K.; Grzesiuk, E.; Poznański, J. Effect of Posttranslational Modifications on the Structure and Activity of FTO Demethylase. Int. J. Mol. Sci. 2021, 22, 4512. https://doi.org/10.3390/ijms22094512

AMA Style

Marcinkowski M, Pilžys T, Garbicz D, Piwowarski J, Mielecki D, Nowaczyk G, Taube M, Gielnik M, Kozak M, Winiewska-Szajewska M, Szołajska E, Dębski J, Maciejewska AM, Przygońska K, Ferenc K, Grzesiuk E, Poznański J. Effect of Posttranslational Modifications on the Structure and Activity of FTO Demethylase. International Journal of Molecular Sciences. 2021; 22(9):4512. https://doi.org/10.3390/ijms22094512

Chicago/Turabian Style

Marcinkowski, Michał, Tomaš Pilžys, Damian Garbicz, Jan Piwowarski, Damian Mielecki, Grzegorz Nowaczyk, Michał Taube, Maciej Gielnik, Maciej Kozak, Maria Winiewska-Szajewska, Ewa Szołajska, Janusz Dębski, Agnieszka M. Maciejewska, Kaja Przygońska, Karolina Ferenc, Elżbieta Grzesiuk, and Jarosław Poznański. 2021. "Effect of Posttranslational Modifications on the Structure and Activity of FTO Demethylase" International Journal of Molecular Sciences 22, no. 9: 4512. https://doi.org/10.3390/ijms22094512

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Article Access Map by Country/Region

1
Back to TopTop