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The energy currency of the cell ATP, is used by kinases to drive key cellular processes. However, the connection of cellular ATP abundance and protein stability is still under investigation. Using Fast Relaxation Imaging paired with alanine scanning and ATP depletion experiments, we study the nucleotide kinase (APSK) domain of 3′-phosphoadenosine-5′-phosphosulfate (PAPS) synthase, a marginally stable protein. Here, we show that the in-cell stability of the APSK is determined by ligand binding and directly connected to cellular ATP levels. The observed protein stability change for different ligand-bound states or under ATP-depleted conditions ranges from ΔGf0 = -10.7 to +13.8 kJ/mol, which is remarkable since it exceeds changes measured previously, for example upon osmotic pressure, cellular stress or differentiation. The results have implications for protein stability during the catalytic cycle of APS kinase and suggest that the cellular ATP level functions as a global regulator of kinase activity.
|Journal||Frontiers in Molecular Bioscience|
|Publication status||Published - 13 Dec 2021|
Bibliographical noteFunding Information:
We acknowledge funding from the Cluster of Excellence RESOLV (EXC 1069). OB was supported by the Graduate School of Solvation Science (Ruhr University Bochum). JM was supported by the European Commission Marie Curie Fellowship SUPA-HD (625451) and the Biochemical Society UK (Eric Reid Fund).
Copyright © 2021 Brylski, Shrestha, Gnutt, Gnutt, Mueller and Ebbinghaus.
- ATP depletion
- PAPS synthase
- alanine scanning
- cellular stress
- in-cell spectroscopy
- ligand binding
- protein folding stability
- sulfation pathways
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
- Molecular Biology
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- 1 Finished
FP7_MC_IEF_SUPA-HD:SULFATION PATHWAYS IN HEALTH AND DISEASE
1/03/14 → 29/02/16