title: Incorporation of bisphosphonates into adenine nucleotides by amoebae of the cellular slime mould Dictyostelium discoideum.
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.
authors.affiliation: Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, U.K.
authors.forename: M J
authors.initials: MJ
authors.lastname: Rogers
authors.forename: X
authors.initials: X
authors.lastname: Ji
authors.forename: R G
authors.initials: RG
authors.lastname: Russell
authors.forename: G M
authors.initials: GM
authors.lastname: Blackburn
authors.forename: M P
authors.initials: MP
authors.lastname: Williamson
authors.forename: A V
authors.initials: AV
authors.lastname: Bayless
authors.forename: F H
authors.initials: FH
authors.lastname: Ebetino
authors.forename: D J
authors.initials: DJ
authors.lastname: Watts
mesh_terms: D000227:Adenine Nucleotides / Q000302:isolation & purification / Q000378:metabolism*; D000604:Amino Acyl-tRNA Synthetases / Q000037:antagonists & inhibitors; D000818:Animals; D002474:Cell-Free System; D002853:Chromatography, Liquid; D004023:Dictyostelium / Q000378:metabolism*; D004164:Diphosphonates / Q000378:metabolism* / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000302:isolation & purification / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D010010:Osteoclasts / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism
chemical_list: D000227:Adenine Nucleotides; D004164:Diphosphonates; D012346:RNA, Transfer, Amino Acyl; D006160:Guanosine Triphosphate; D000604:Amino Acyl-tRNA Synthetases