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Purification and biochemical characterization of acetylcholinesterase (AChE) from the excretory/secretory products of Trichostrongylus colubriformis

Published online by Cambridge University Press:  06 April 2009

G. Griffiths  and
D. I. Pritchard
G. Griffiths
Affiliation:
Department of Life Science, University of Nottingham, University Park, Nottingham NG7 2RD
D. I. Pritchard
Affiliation:
Department of Life Science, University of Nottingham, University Park, Nottingham NG7 2RD
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Summary

Acetylcholinesterase (AChE) has been purified from the excretory/secretory (ES) products of Trichostrongylus colubriformis (using edrophonium chloride linked to epoxy-activated Sepharose) with yields of 40–50%. Purity was confirmed by polyacrylamide gel electrophoresis (using silver [protein] and Karnovsky [activity] stains) and measurement of specific AChE activity. Further analysis of the purified fractions by gel filtration and sucrose density gradient techniques revealed the existence of 2 forms of hydrophilic AChE (Mr 189 and 80 kDa). From the data we deduce these to be the globular monomer and dimer, G1 and G2 forms of AChE. Inhibition studies using BW284C51, iso-OMPA and excess substrate, along with substrate specificity studies, show both forms to be true acetylcholinesterases. We are currently assessing the protective immunogenicity of purified AChE in sheep.

Keywords

acetylcholinesterasepurificationTrichostrongylus colubriformismolecular forms
Type
Research Article
Information
Parasitology , Volume 108 , Issue 5 , June 1994 , pp. 579 - 586
Copyright
Copyright © Cambridge University Press 1994

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