Korean J Parasitol.  2004 Jun;42(2):57-60. 10.3347/kjp.2004.42.2.57.

Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

• Chung YB
• Kong Y
• Yang HJ

• KMID: 1051419
• DOI: http://doi.org/10.3347/kjp.2004.42.2.57

Abstract

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

Keyword

sparganum; antigenicity; N-glycosylation; ELISA

MeSH Terms

Animals
Antigens, Helminth/*analysis/chemistry/immunology/metabolism
Carbohydrates/analysis/immunology
Chromatography, Affinity
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Epitopes/analysis/immunology
Glucosaminidase/metabolism
Human
Peptide-N4- (N-acetyl-beta-glucosaminyl) Asparagine Amidase/metabolism
Periodic Acid/chemistry
Sparganosis/*parasitology
Sparganum/immunology/metabolism
Spirometra/immunology/*metabolism
Support, Non-U.S. Gov't