Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum

Michael John Rogers, X Ji, R G Russell, G M Blackburn, M P Williamson, A V Bayless, F H Ebetino, D J Watts

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Bisphosphonates are a class of synthetic pyrophosphate analogues. Some are known to be potent inhibitors of osteoclast-mediated bone resorption in vivo, but their mechanisms of action are unclear. The order of potency of bisphosphonates as inhibitors of bone resorption closely matches the order of potency as inhibitors of growth of amoebae of the slime mould Dictyostelium discoideum, indicating that bisphosphonates may have a mechanism of action that is similar in both osteoclasts and Dictyostelium. Methylenebisphosphonate and several halogenated derivatives, which have low potency as antiresorptive agents and as growth inhibitors of Dictyostelium, are metabolized intracellularly by Dictyostelium amoebae into methylene-containing adenine nucleotides. We have used a combination of n.m.r. and f.p.l.c. analysis to determine whether incorporation into nucleotides is a feature of other bisphosphonates, especially those that are potent antiresorptive agents. Only bisphosphonates with short side chains or of low potency are incorporated into adenine nucleotides, whereas those with long side chains or of high potency are not metabolized. Bisphosphonate metabolism in cell-free extracts of Dictyostelium was accompanied by inhibition of aminoacylation of tRNA by several aminoacyl-tRNA synthetases. These enzymes were barely affected by the bisphosphonates that were not metabolized. The results indicate that some bisphosphonates are not metabolically inert analogues of pyrophosphate and appear to be metabolized by aminoacyl-tRNA synthetases. The cellular effects of some bisphosphonates may be the result of their incorporation into adenine nucleotides or inhibition of aminoacyl-tRNA synthetases, although the potent bisphosphonates appear to act by a different mechanism.
Original languageEnglish
Pages (from-to)303-11
Number of pages9
JournalBiochemical Journal
Volume303 ( Pt 1)
Publication statusPublished - 1 Oct 1994

Fingerprint

Dictyosteliida
Amoeba
Dictyostelium
Adenine Nucleotides
Diphosphonates
Bone Density Conservation Agents
Amino Acyl-tRNA Synthetases
Growth Inhibitors
Osteoclasts
Transfer RNA Aminoacylation
Bone Resorption
Transfer RNA
Cell Extracts
Metabolism
Bone
Fungi
Nucleotides

Keywords

  • Adenine Nucleotides
  • Amino Acyl-tRNA Synthetases
  • Animals
  • Cell-Free System
  • Chromatography, Liquid
  • Dictyostelium
  • Diphosphonates
  • Guanosine Triphosphate
  • Magnetic Resonance Spectroscopy
  • Osteoclasts
  • RNA, Transfer, Amino Acyl

Cite this

Rogers, M. J., Ji, X., Russell, R. G., Blackburn, G. M., Williamson, M. P., Bayless, A. V., ... Watts, D. J. (1994). Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum. Biochemical Journal, 303 ( Pt 1), 303-11.

Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum. / Rogers, Michael John; Ji, X; Russell, R G; Blackburn, G M; Williamson, M P; Bayless, A V; Ebetino, F H; Watts, D J.

In: Biochemical Journal, Vol. 303 ( Pt 1), 01.10.1994, p. 303-11.

Research output: Contribution to journalArticle

Rogers, MJ, Ji, X, Russell, RG, Blackburn, GM, Williamson, MP, Bayless, AV, Ebetino, FH & Watts, DJ 1994, 'Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum', Biochemical Journal, vol. 303 ( Pt 1), pp. 303-11.
Rogers MJ, Ji X, Russell RG, Blackburn GM, Williamson MP, Bayless AV et al. Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum. Biochemical Journal. 1994 Oct 1;303 ( Pt 1):303-11.
Rogers, Michael John ; Ji, X ; Russell, R G ; Blackburn, G M ; Williamson, M P ; Bayless, A V ; Ebetino, F H ; Watts, D J. / Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum. In: Biochemical Journal. 1994 ; Vol. 303 ( Pt 1). pp. 303-11.
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abstract = "Bisphosphonates are a class of synthetic pyrophosphate analogues. Some are known to be potent inhibitors of osteoclast-mediated bone resorption in vivo, but their mechanisms of action are unclear. The order of potency of bisphosphonates as inhibitors of bone resorption closely matches the order of potency as inhibitors of growth of amoebae of the slime mould Dictyostelium discoideum, indicating that bisphosphonates may have a mechanism of action that is similar in both osteoclasts and Dictyostelium. Methylenebisphosphonate and several halogenated derivatives, which have low potency as antiresorptive agents and as growth inhibitors of Dictyostelium, are metabolized intracellularly by Dictyostelium amoebae into methylene-containing adenine nucleotides. We have used a combination of n.m.r. and f.p.l.c. analysis to determine whether incorporation into nucleotides is a feature of other bisphosphonates, especially those that are potent antiresorptive agents. Only bisphosphonates with short side chains or of low potency are incorporated into adenine nucleotides, whereas those with long side chains or of high potency are not metabolized. Bisphosphonate metabolism in cell-free extracts of Dictyostelium was accompanied by inhibition of aminoacylation of tRNA by several aminoacyl-tRNA synthetases. These enzymes were barely affected by the bisphosphonates that were not metabolized. The results indicate that some bisphosphonates are not metabolically inert analogues of pyrophosphate and appear to be metabolized by aminoacyl-tRNA synthetases. The cellular effects of some bisphosphonates may be the result of their incorporation into adenine nucleotides or inhibition of aminoacyl-tRNA synthetases, although the potent bisphosphonates appear to act by a different mechanism.",
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N2 - Bisphosphonates are a class of synthetic pyrophosphate analogues. Some are known to be potent inhibitors of osteoclast-mediated bone resorption in vivo, but their mechanisms of action are unclear. The order of potency of bisphosphonates as inhibitors of bone resorption closely matches the order of potency as inhibitors of growth of amoebae of the slime mould Dictyostelium discoideum, indicating that bisphosphonates may have a mechanism of action that is similar in both osteoclasts and Dictyostelium. Methylenebisphosphonate and several halogenated derivatives, which have low potency as antiresorptive agents and as growth inhibitors of Dictyostelium, are metabolized intracellularly by Dictyostelium amoebae into methylene-containing adenine nucleotides. We have used a combination of n.m.r. and f.p.l.c. analysis to determine whether incorporation into nucleotides is a feature of other bisphosphonates, especially those that are potent antiresorptive agents. Only bisphosphonates with short side chains or of low potency are incorporated into adenine nucleotides, whereas those with long side chains or of high potency are not metabolized. Bisphosphonate metabolism in cell-free extracts of Dictyostelium was accompanied by inhibition of aminoacylation of tRNA by several aminoacyl-tRNA synthetases. These enzymes were barely affected by the bisphosphonates that were not metabolized. The results indicate that some bisphosphonates are not metabolically inert analogues of pyrophosphate and appear to be metabolized by aminoacyl-tRNA synthetases. The cellular effects of some bisphosphonates may be the result of their incorporation into adenine nucleotides or inhibition of aminoacyl-tRNA synthetases, although the potent bisphosphonates appear to act by a different mechanism.

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