Characterisation of the secretory acetylcholinesterases from adult Nippostrongylus brasiliensis

C. Clare Blackburn*, Murray E. Selkirk

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

The biochemical nature and relationship between the different isoforms of acetylcholinesterase (AChEs) secreted by adult Nippostrongylus brasiliensis was investigated, primarily via staining for enzyme activity and active-site labelling with [3H]-diisopropylfluorophosphate (DFP). Analysis by 1-dimensional SDS-PAGE under non-reducing conditions revealed the existence of 2 proteins of 74-kDa and 39-kDa, and each protein resolved as 2 species by isoelectric focussing. Both AChEs were co-purified via affinity chromatography on 9-[Nβ-(ε{lunate}-aminocaproyl)-β-aminopropylam ino]-acridine-coupled Sepharose 6B, and utilised to raise a polyclonal rabbit antiserum. Examination of the expression of secretory AChEs by adult worms during their residence in the gastrointestinal tract showed that the initial secretion of both forms on day 4 post-infection switched to predominant secretion of the 39-kDa protein by day 8. Immunoprecipitation of 35S-labelled products of in vitro translation via RNA from day 4 and day 8 worms predicted a single primary translation product of 59 kDa. These data suggested that the 'switching' event seen in vivo most likely corresponded to processing of the 74-kDa molecule. This interpretation was supported by limited digestion with V8 protease and chymotrypsin, which showed that the 74-kDa and the 39-kDa proteins possessed structural similarities.

Original languageEnglish
Pages (from-to)79-88
Number of pages10
JournalMolecular and Biochemical Parasitology
Issue number1-2
Publication statusPublished - 1 Jan 1992

Keywords / Materials (for Non-textual outputs)

  • Acetylcholinesterase
  • Excretory/secretory products
  • Nippostrongylus brasiliensis


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