Cellular ATP levels determine the stability of a nucleotide kinase

Oliver Brylski, Puja Shrestha, Patricia Gnutt, David Gnutt, Jonathan Wolf Mueller, Simon Ebbinghaus

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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.
Original languageEnglish
Article number790304
JournalFrontiers in Molecular Bioscience
Publication statusPublished - 13 Dec 2021

Bibliographical note

Funding 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).

Publisher Copyright:
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
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Molecular Biology


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