title: Farsenyl pyrophosphate synthase is a potential molecular drug target of risedronate in Babesia bovis.
abstract: A cDNA encoding farnesyl pyrophosphate synthase of Babesia bovis (BbFPPS) has been isolated, cloned and characterized as molecular drug target. Sequence analysis revealed that BbFPPS contains an open reading frame of 1011bp with predicted 336 amino acids and molecular mass of 38kDa. Antiserum raised in mice against recombinant BbFPPS expressed in Escherichia coli specifically reacted with native protein of B. bovis parasites by Western blot analysis and indirect immunofluorescent test. Enzymatic assay using recombinant BbFPPS revealed that the Km value of the enzyme for isopentenyl pyrophosphate and dimethylallyl pyrophosphate was 2.494±1.536μM. Risedronate inhibited the activity of BbFPPS yielding IC50 value of 8.4±1.2nM. Furthermore, the in vitro growth of B. bovis was significantly inhibited in the presence of a micromolar concentration of risedronate (IC50=4.02±0.91μM). No regrowth of B. bovis was observed at 10μM of risedronate in the subsequent viability test. These results demonstrate that BbFPPS is the molecular target of risedronate, which could inhibit the in vitro growth of B. bovis.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 2-13 Inada-cho, Obihiro, Hokkaido, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Ueno
authors.forename: Mohamad Alaa
authors.initials: MA
authors.lastname: Terkawi
authors.forename: Miki
authors.initials: M
authors.lastname: Yokoyama
authors.forename: Shinuo
authors.initials: S
authors.lastname: Cao
authors.forename: Gabriel
authors.initials: G
authors.lastname: Aboge
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Aboulaila
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Nishikawa
authors.forename: Xuenan
authors.initials: X
authors.lastname: Xuan
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.forename: Ikuo
authors.initials: I
authors.lastname: Igarashi
mesh_terms: D000818:Animals; D016793:Babesia bovis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D012968:Etidronic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D051228:Geranyltranstransferase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D045782:Hemiterpenes / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D009943:Organophosphorus Compounds / Q000378:metabolism; D018512:Parasitemia; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000068296:Risedronic Acid; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D016054:DNA, Protozoan; D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D015800:Protozoan Proteins; C004809:isopentenyl pyrophosphate; C043060:3,3-dimethylallyl pyrophosphate; D051228:Geranyltranstransferase; D000068296:Risedronic Acid; D012968:Etidronic Acid