Structural analysis of the glycoprotein allergen Hev b 4 from natural rubber latex by mass spectrometry

Daniel Kolarich, Friedrich Altmann*, Elumalai Sunderasan

*Corresponding author for this work

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

The lecithinase homolog (Hev b 4) from Hevea brasiliensis (Q6T4P0_HEVBR) is an important natural rubber latex allergen. Hev b 4 is a highly glycosylated protein and its carbohydrate moiety has been implicated in the binding of IgE from natural rubber latex allergic patients. The cDNA for Hev b 4 has recently been cloned and sequenced. Here, we have analyzed the post-translational modifications of natural Hev b 4 by liquid chromatography/electrospray ionization-mass spectrometry of tryptic peptides. Seven of the eight potential glycosylation sites were found to be occupied. One site, however, was only partially glycosylated. Asn224 was substituted by complex type N-glycans with fucose and xylose, whereas all other sites carried either oligomannose glycans or a mixture of oligomannose and complex N-glycans. Glycosylation site Asn308, the most C-terminal one of the eight sites, was only found in the non-glycosylated form. The complex type N-glycans apparently form the molecular basis for the immune reaction with patients' sera. A large fraction of Hev b 4 molecules contains two or more complex N-glycans and thus a physiological reaction against these polyvalent allergens on the basis of the carbohydrate is in theory possible. Aside from allowing glycosylation analysis, the mass spectrometric data defined the N-terminal cleavage site of Hev b 4. This study once more demonstrates the outstanding analytical potential of electrospray ionization-mass spectrometry coupled with liquid chromatographic separation.

Original languageEnglish
Pages (from-to)715-720
Number of pages6
JournalBiochimica et Biophysica Acta - General Subjects
Volume1760
Issue number4
DOIs
Publication statusPublished - Apr 2006

Keywords

  • ESI-MS
  • Glycoprotein
  • Glycoproteomics
  • Hevea brasiliensis
  • Latex allergy
  • Natural rubber latex

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