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Variation in the nature of attachment of phosphorylcholine to excretory–secretory products of adult Brugia pahangi

Published online by Cambridge University Press:  01 March 1997

Z. M. NOR
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR
K. M. HOUSTON
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR
E. DEVANEY
Affiliation:
Department of Veterinary Parasitology, University of Glasgow, G61 1GH
W. HARNETT
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR

Abstract

The mechanism of linkage of phosphorylcholine (PC) to excretory–secretory products (ES) of adult Brugia pahangi has been investigated. Biosynthetic radio-isotope labelling of ES with [3H]choline followed by SDS–PAGE/fluorography revealed a smear of molecular weight approximately 40–100 kDa which loses its radiolabel following exposure to N-glycosidase F, but not mild alkali. PC is thus attached to this smear of molecules via N-type glycans, a mechanism of linkage previously observed with respect to PC–ES of Acanthocheilonema viteae. Western blotting analysis of non-radiolabelled ES demonstrated the existence of additional PC–ES which were insensitive to N-glycosidase F, but not to alkali. This second group of molecules is therefore likely to contain PC linked to O-glycans. Filarial nematodes may thus utilize 2 classes of glycan for attachment of PC. Examination of B. pahangi and A. viteae whole worm extracts by Western blotting indicated that their PC content could not be cleaved by N-glycosidase F and hence the use of N-type glycans may be restricted to a subset of ES products. The implications of these findings with respect to developing inhibitors of PC attachment for use as anti-filarial drugs are discussed.

Type
Research Article
Copyright
1997 Cambridge University Press

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