title: A distinct family of acetylcholinesterases is secreted by Nippostrongylus brasiliensis.
abstract: A third variant of acetylcholinesterase (AChE A) secreted by the parasitic nematode Nippostrongylus brasiliensis has been isolated which shows 63-64% identity to AChE B and AChE C, with a truncated carboxyl terminus and a short internal insertion relative to AChEs from other species. Three of the fourteen aromatic residues which line the active site gorge in Torpedo AChE are substituted by non-aromatic residues (Y70T, W279D and F288M). All three enzymes have 8 cysteine residues in conserved positions, including 6 which have been implicated in disulphide bonds in other AChEs. Phylogenetic analysis suggests that these enzymes form a distinct group which evolved after speciation and are most closely related to ACE-2 of Caenorhabditis elegans. Recombinant AChE A secreted by Pichia pastoris was monomeric and hydrophilic, with a substrate preference for acetylthiocholine and negligible activity against butyrylthiocholine. A model structure of AChE A built from the coordinates of the Torpedo californica AChE suggests that W345 (F331 in Torpedo) limits the docking of butyrylcholine. This model is consistent with mutational analysis of the nematode enzymes. Expression of AChE A is regulated at the transcriptional level independently of the other 2 secreted variants, with maximal expression by fourth stage larvae and young adult worms. These enzymes thus appear to represent an unusual family of AChEs with conserved structural features which operate outside the normal boundaries of known functions in regulation of endogenous neurotransmitter activity.
authors.affiliation: Department of Biological Sciences, Biochemistry Building, Imperial College of Science, Technology and Medicine, South Kensington Campus, London SW7 2AY, UK.
authors.forename: Ayman S
authors.initials: AS
authors.lastname: Hussein
authors.forename: Michal
authors.initials: M
authors.lastname: Harel
authors.forename: Murray E
authors.initials: ME
authors.lastname: Selkirk
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000122:Acetylthiocholine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002092:Butyrylthiocholine / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009559:Nippostrongylus / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D010843:Pichia / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D000122:Acetylthiocholine; D002092:Butyrylthiocholine; D000110:Acetylcholinesterase