The N-glycans of yellow jacket venom hyaluronidases and the protein sequence of its major isoform in Vespula vulgaris.
Hyaluronidase (E.C. 3.2.1.35), one of the three major allergens of yellow jacket venom, is a glycoprotein of 45 kDa that is largely responsible for the cross-reactivity of wasp and bee venoms with sera of allergic patients. The asparagine-linked carbohydrate often appears to constitute the common IgE-binding determinant. Using a combination of MALDI MS and HPLC of 2-aminopyridine-labelled glycans, we found core-difucosylated paucimannosidic glycans to be the major species in the 43-45 kDa band of Vespula vulgaris and also in the corresponding bands of venoms from five other wasp species (V. germanica, V. maculifrons, V. pensylvanica, V. flavopilosa and V. squamosa). Concomitant peptide mapping of the V. vulgaris 43 kDa band identified the known hyaluronidase, Ves v 2 (SwissProt P49370), but only as a minor component. De novo sequencing by tandem MS revealed the predominating peptides to resemble a different, yet homologous, sequence. cDNA cloning retrieved a sequence with 58 and 59% homology to the previously known isoform and to the Dolichovespula maculata and Polistes annularis hyaluronidases. Close homologues of this new, putative hyaluronidase b (Ves v 2b) were also the major isoform in the other wasp venoms.
Kolarich
Daniel
D
Department of Chemistry, University of Natural Resources and Applied Life Sciences (BOKU), Vienna, Austria.
Léonard
Renaud
R
Hemmer
Wolfgang
W
Altmann
Friedrich
F
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D006821:Hyaluronoglucosaminidase / Q000737:chemistry / Q000235:genetics*; D006927:Hymenoptera / Q000201:enzymology / Q000235:genetics; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D011134:Polysaccharides / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014862:Wasp Venoms / Q000737:chemistry / Q000201:enzymology*; D014863:Wasps / Q000201:enzymology* / Q000235:genetics
D007527:Isoenzymes; D011134:Polysaccharides; D014862:Wasp Venoms; D006821:Hyaluronoglucosaminidase