Fluoride exposure inhibits protein expression and enzyme activity in the lung-stage larvae of Ascaris suum.
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.
Islam
M K
MK
Laboratory of Parasitic Diseases, National Institute of Animal Health, National Agricultural Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.
Miyoshi
T
T
Yamada
M
M
Alim
M A
MA
Huang
X
X
Motobu
M
M
Tsuji
N
N
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
D015801:Helminth Proteins; D012969:Sodium Fluoride; D011755:Pyrophosphatases