Expression of a single dimeric membrane-bound acetylcholinesterase in Parascaris equorum.
A single form of cholinesterase was detected in the parasitic nematode Parascaris equorum and purified from a low-salt Triton X-100 extract of whole animals by affinity chromatography on an edrophonium-Sepharose matrix. Based on gel-filtration chromatography, sedimentation analysis and SDS-PAGE, such a cholinesterase is an amphiphilic globular (G2) dimer (125-129 kDa, 6.1 S). It includes some hydrophobic domain other than phosphatidylinositol, which gives autoaggregation, detergent interaction and also anchors the molecule to the cell membrane. The enzyme, probably functional in cholinergic neurotransmission, is an acetylcholinesterase showing a fairly low substrate specificity with thiocholine esters. Electrostatic interactions seem to play a major role in the catalytic activity. Studies with inhibitors gave complete inhibition with 1 mM eserine, low sensitivity for procainamide and for tetra(monoisopropyl)pyrophosphortetramide as well as higher inhibition with edrophonium chloride and 1,5-bis(4allyldimethylammoniumphenyl)-pentan-3-one dibromide. The enzyme also showed excess-substrate inhibition with acetylthiocholine. No cross-hybridization occurred between the gene(s) encoding acetylcholinesterase in P. equorum and ace-1 from the free-living nematode Caenorhabditis elegans. The expression of a single cholinesterase form in P. equorum, unusual in free-living nematodes, could be due to parasitic life adaptation with resulting reduction of locomotor activity.
Talesa
V
V
Department of Experimental Medicine, University of Perugia, Italy.
Romani
R
R
Grauso
M
M
Rosi
G
G
Giovannini
E
E
D000110:Acetylcholinesterase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001199:Ascaridoidea / Q000201:enzymology*; D015139:Blotting, Southern; D002499:Centrifugation, Density Gradient; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D017238:Genes, Helminth; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000378:metabolism
D002800:Cholinesterase Inhibitors; D000110:Acetylcholinesterase; D010738:Type C Phospholipases