Characterization of secretory acetylcholinesterase from Setaria cervi microfilariae: a potential antigen for diagnosis of human filariasis.
Acetylcholinesterase (AChE) is released to the external medium when microfilariae (m.f.) of Setaria cervi, a bovine filarial parasite, are maintained in vitro. Intense enzyme staining at amphids, excretory pores, anal vesicle and phasmids suggest an active secretion of AChE from m.f. Excretory-secretory products of m.f. displayed two electromorphic variants of AChE when resolved by 6% nondenaturing PAGE. The two isoforms of AChE (A and B) were separated on the basis of charge by DEAE sepharose CL 6B column following gel filtration. The two isoforms showed differing kinetic properties with respect to substrate specificity and inhibitor sensitivity. Anti-Nippostrongylus brasiliensis AChE antibodies cross-reacted with the affinity purified secretory AChE in ELISA. Immunoblotting of purified AChEs with cross-reacting anti-AChE antibodies revealed the presence of an approximately 75 kD protein in the isoenzyme A and an approximately 45 kD protein in B, whereas both proteins were present in the enzyme purified via affinity chromatography on edrophonium sepharose column.
Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi, India.
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Sharma
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Rathaur
D000110:Acetylcholinesterase / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000947:Antigens, Helminth; D002845:Chromatography; D004797:Enzyme-Linked Immunosorbent Assay; D005368:Filariasis / Q000175:diagnosis* / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D017182:Setaria Nematode / Q000201:enzymology* / Q000276:immunology
D000947:Antigens, Helminth; D000110:Acetylcholinesterase