Structural analysis of the glycoprotein allergen Art v II from the pollen of mugwort (Artemisia vulgaris L.)

J Biol Chem. 1991 Feb 5;266(4):2660-8.

Abstract

The glycoprotein allergen Art v II, from the pollen of mugwort (Artemisia vulgaris L.) was treated with peptide:N-glycosidase F (PNGase F) to release asparagine-linked oligosaccharides. The oligosaccharides were isolated by gel permeation chromatography and their structures determined by 500-MHz 1H NMR spectroscopy, fast atom bombardment-mass spectrometry, and high-pH anion-exchange chromatography. The high-mannose oligosaccharides Man5GlcNAc2, Man6GlcNAc2, Man7GlcNAc2, Man8GlcNAc2, and Man9GlcNAc2 were present in the ratios 2:49:19:24:6 and accounted for all the asparagine-linked oligosaccharides released from Art v II by PNGase F. The NH2-terminal amino acid sequences of Art v II and of four peptides generated by cyanogen bromide (CNBr) cleavage of deglycosylated Art v II were determined. The first 30 amino acid residues of Art v II did not contain any potential N-glycosylation sites. One potential N-glycosylation site was identified in one of the CNBr fragments. The native protein conformation was shown by enzyme-linked immunosorbent assay inhibition assays to be essential for the binding of rabbit IgG to Art v II and for the binding of human IgE to the major IgE-binding epitope(s) in this allergen. At least one minor IgE-binding epitope still bound IgE after denaturation of the allergen. Removal of the high-mannose chains from denatured Art v II had no significant effect on the binding of human IgE to the minor IgE-binding epitope(s).

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Allergens*
  • Amino Acid Sequence
  • Animals
  • Antigens, Plant
  • Asparagine / metabolism
  • Carbohydrate Sequence
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Cyanogen Bromide / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes
  • Glycoproteins / chemistry*
  • Glycoproteins / immunology
  • Glycoside Hydrolases / metabolism
  • Immunoglobulin G / immunology
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Oligosaccharides / chemistry
  • Plant Proteins*
  • Plants*
  • Pollen* / immunology
  • Protein Conformation
  • Rabbits

Substances

  • Allergens
  • Antigens, Plant
  • Epitopes
  • Glycoproteins
  • Immunoglobulin G
  • Oligosaccharides
  • Plant Proteins
  • allergen Art v II protein, Artemisia vulgaris L.
  • Asparagine
  • Glycoside Hydrolases
  • Cyanogen Bromide