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

Parasitology. 1997 Mar:114 ( Pt 3):257-62. doi: 10.1017/s0031182096008402.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / metabolism
  • Animals
  • Antigens, Helminth / chemistry
  • Antigens, Helminth / metabolism*
  • Blotting, Western
  • Brugia pahangi / chemistry*
  • Brugia pahangi / immunology
  • Electrophoresis, Polyacrylamide Gel
  • Helminth Proteins / chemistry
  • Helminth Proteins / metabolism*
  • Molecular Weight
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
  • Phosphorylcholine / chemistry
  • Phosphorylcholine / metabolism*
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism*

Substances

  • Antigens, Helminth
  • Helminth Proteins
  • Polysaccharides
  • Phosphorylcholine
  • Amidohydrolases
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase