Cellular ATP levels determine the stability of a nucleotide kinase

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

Research output: Contribution to journalArticlepeer-review

Abstract

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
Volume8
DOIs
Publication statusPublished - 13 Dec 2021

Keywords

  • PAPS synthase
  • ATP depletion
  • in-cell spectroscopy
  • protein folding stability
  • alanine scanning
  • ligand binding
  • sulfation pathways
  • cellular stress

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