title: Fluoride exposure inhibits protein expression and enzyme activity in the lung-stage larvae of Ascaris suum.
abstract: Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process of Ascaris suum larvae. This study describes moulting and development-specific protein expression profiles of A. suum lung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3-hydroxyacyl-CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.
authors.affiliation: Laboratory of Parasitic Diseases, National Institute of Animal Health, National Agricultural Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.
authors.forename: M K
authors.initials: MK
authors.lastname: Islam
authors.forename: T
authors.initials: T
authors.lastname: Miyoshi
authors.forename: M
authors.initials: M
authors.lastname: Yamada
authors.forename: M A
authors.initials: MA
authors.lastname: Alim
authors.forename: X
authors.initials: X
authors.lastname: Huang
authors.forename: M
authors.initials: M
authors.lastname: Motobu
authors.forename: N
authors.initials: N
authors.lastname: Tsuji
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017165:Ascaris suum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods / Q000662:veterinary; D005786:Gene Expression Regulation; D015801:Helminth Proteins / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D007814:Larva; D008168:Lung / Q000469:parasitology; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence / Q000662:veterinary; D018978:Molting / Q000187:drug effects*; D011755:Pyrophosphatases / Q000187:drug effects* / Q000378:metabolism; D011817:Rabbits; D012969:Sodium Fluoride / Q000494:pharmacology*; D013047:Specific Pathogen-Free Organisms
chemical_list: D015801:Helminth Proteins; D012969:Sodium Fluoride; D011755:Pyrophosphatases