Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation.
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.
Chung
Young-Bae
YB
Department of Parasitology, College of Medicine and Institute of Medicine, Cheju National University, Jeju 690-756, Korea. parayang@ewha.ac.kr
Kong
Yoon
Y
Yang
Hyun-Jong
HJ
D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis / Q000276:immunology; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D010504:Periodic Acid / Q000737:chemistry; D013031:Sparganosis / Q000469:parasitology*; D013032:Sparganum / Q000276:immunology / Q000378:metabolism; D013146:Spirometra / Q000276:immunology / Q000378:metabolism*
D000947:Antigens, Helminth; D002241:Carbohydrates; D000939:Epitopes; D010504:Periodic Acid; C009288:metaperiodate; D006596:Hexosaminidases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase