A phosphofructokinase homolog from Pyrobaculum calidifontis displays kinase activity towards pyrimidine nucleosides and ribose 1-phosphate

Iram Aziz, Tahira Bibi, Naeem Rashid*, Riku Aono, Haruyuki Atomi, Muhammad Akhtar

*この研究の対応する著者

研究成果査読

5 被引用数 (Scopus)

抄録

The genome of the hyperthermophilic archaeon Pyrobaculum calidifontis contains an open reading frame, Pcal_0041, annotated as encoding a PfkB family ribokinase, consisting of phosphofructokinase and pyrimidine kinase domains. Among the biochemically characterized enzymes, the Pcal_0041 protein was 37% identical to the phosphofructokinase (Ape_0012) from Aeropyrum pernix, which displayed kinase activity toward a broad spectrum of substrates, including sugars, sugar phosphates, and nucleosides, and 36% identical to a phosphofructokinase from Desulfurococcus amylolyticus. To examine the biochemical function of the Pcal_0041 protein, we cloned and expressed the gene and purified the recombinant protein. Although the Pcal_0041 protein contained a putative phosphofructokinase domain, it exhibited only low levels of phosphofructokinase activity. The recombinant enzyme catalyzed the phosphorylation of nucleosides and, to a lower extent, sugars and sugar phosphates. Surprisingly, among the substrates tested, the highest activity was detected with ribose 1-phosphate (R1P), followed by cytidine and uridine. The catalytic efficiency (k cat /K m ) toward R1P was 11.5 mM -1 · s -1 . ATP was the most preferred phosphate donor, followed by GTP. Activity measurements with cell extracts of P. calidifontis indicated the presence of nucleoside phosphorylase activity, which would provide the means to generate R1P from nucleosides. The study suggests that, in addition to the recently identified ADP-dependent ribose 1-phosphate kinase (R1P kinase) in Thermococcus kodakarensis that functions in the pentose bisphosphate pathway, R1P kinase is also present in members of the Crenarchaeota.

本文言語English
論文番号e00284-18
ジャーナルJournal of Bacteriology
200
16
DOI
出版ステータスPublished - 2018 8 1

ASJC Scopus subject areas

  • 微生物学
  • 分子生物学

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