Pritchard, D. I. and Leggett, K. V. and Rogan, M. T. and McKean, Paul G. and Brown, Alan (1991) Necator americanus secretory acetylcholinesterase and its purification from excretory-secretory products by affinity chromatography. Parasite Immunology, 13 (2). pp. 187-199. ISSN 1365-3024Full text not available from this repository.
Acetylcholinesterase (AChE) secretion by adult N. americanus was enhanced in vitro by incorporating insoluble collagen rafts into culture dishes. Enzyme produced in this way had preferential substrate specificity for acetylthiocholine iodide (ATC), and its activity was inhibited by eserine (1.1 x 10(-8) M). Ancylostoma ceylanicum, another hookworm species, failed to produce comparable amounts of AChE in culture. AChE was efficiently purified from culture medium by affinity chromatography on edrophonium sepharose; 81% of the AChE activity was retained by the affinity matrix, although this fraction contained only 4.3% of the protein loaded. Antisera raised against purified AChE in rabbits immunohistochemically stained the oesophageal glands of the parasite, and reacted with molecules of 32, 60, 80, 140 and 220 kDa in reduced adult ES products on Western blotting, although differential activity was observed against worm homogenates and earlier developmental stages. On IEF, purified AChE resolved predominantly with a pl of 3.55; proteins with a similar pl were recognized by rabbit anti-AChE. IgG preparations of this antiserum inhibited AChE activity in ES products, and inhibited AChE secretion by adult worms in culture. The availability of this immunological probe will allow definitive experiments to be conducted on the role of this enigmatic enzyme in the host-parasite relationship.
|Journal or Publication Title:||Parasite Immunology|
|Uncontrolled Keywords:||AChE • purification • edrophonium-sepharose • IEF • N. americanus • Western blot • immunohistochemistry|
|Subjects:||Q Science > QH Natural history > QH301 Biology|
|Departments:||Faculty of Health and Medicine > Biomedical & Life Sciences|
|Deposited By:||Dr Paul G McKean|
|Deposited On:||09 Mar 2009 16:04|
|Last Modified:||24 Jun 2016 01:17|
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