The kinetic characteristics of human and trypanosomatid phosphofructokinases for the reverse reaction

Peter M Fernandes, James R Kinkead, Iain W McNae, Paul A Michels, Malcolm D Walkinshaw

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Eukaryotic ATP-dependent phosphofructokinases (PFKs) are often considered unidirectional enzymes catalysing the transfer of a phospho moiety from ATP to fructose 6-phosphate (F6P) to produce ADP and fructose 1,6-bisphosphate (F16BP). The reverse reaction is not generally considered to occur under normal conditions and has never been demonstrated for any eukaryotic ATP-dependent PFKs, though it does occur in PPi-dependent PFKs and has been experimentally shown for bacterial ATP-dependent PFKs. Evidence is provided via two orthogonal assays that all three human PFK isoforms can catalyse the reverse reaction in vitro , allowing determination of kinetic properties. Additionally, the reverse reaction was shown possible for PFKs from three clinically important trypanosomatids; these enzymes are contained within glycosomes in vivo This compartmentalisation may facilitate reversal, given the potential for trypanosomatids to have an altered ATP/ADP ratio in glycosomes compared to the cytosol. The kinetic properties of each trypanosomatid PFK were determined, including the response to natural and artificial modulators of enzyme activity. The possible physiological relevance of the reverse reaction in trypanosomatid and human PFKs is discussed.

Original languageEnglish
Pages (from-to)179-191
Number of pages13
JournalBiochemical Journal
Issue number2
Early online date7 Nov 2018
Publication statusPublished - 18 Jan 2019

Keywords / Materials (for Non-textual outputs)

  • Phosphofructokinase
  • Human
  • Trypanosoma cruzi
  • Trypanosoma brucei
  • Leishmania
  • Trypanosomatid
  • Reverse reaction


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