Back to Search Start Over

Structure of recombinant Ves v 2 at 2.0 Å resolution: structural analysis of an allergenic hyaluronidase from wasp venom.

Authors :
Skov, Lars K.
Seppälä, Ulla
Coen, Jeremy J. F.
Crickmore, Neil
King, Te P.
Monsalve, Rafael
Kastrup, Jette S.
Spangfort, Michael D.
Gajhede, Michael
Source :
Acta Crystallographica: Section D (Wiley-Blackwell); Jun2006, Vol. 62 Issue 6, p595-604, 10p, 1 Color Photograph, 7 Diagrams, 1 Chart, 2 Graphs
Publication Year :
2006

Abstract

Wasp venom from Vespula vulgaris contains three major allergens: Ves v 1, Ves v 2 and Ves v 5. Here, the cloning, expression, biochemical characterization and crystal structure determination of the hyaluronidase Ves v 2 from family 56 of the glycoside hydrolases are reported. The allergen was expressed in Escherichia coli as an insoluble protein and refolded and purified to obtain full enzymatic activity. Three N-glycosylation sites at Asn79, Asn99 and Asn127 were identified in Ves v 2 from a natural source by enzymatic digestions combined with MALDI–TOF mass spectrometry. The crystal structure of recombinant Ves v 2 was determined at 2.0 Å resolution and reveals a central (β/α)<subscript>7</subscript> core that is further stabilized by two disulfide bonds (Cys19–Cys308 and Cys185–Cys197). Based on sequence alignments and structural comparison with the honeybee allergen Api m 2, it is proposed that a conserved cavity near the active site is involved in binding of the substrate. Surface epitopes and putative glycosylation sites have been compared with those of two other major group 2 allergens from Apis mellifera (honeybee) and Dolichovespula maculata (white-faced hornet). The analysis suggests that the harboured allergic IgE-mediated cross-reactivity between Ves v 2 and the allergen from D. maculata is much higher than that between Ves v 2 and the allergen from A. mellifera. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
09074449
Volume :
62
Issue :
6
Database :
Complementary Index
Journal :
Acta Crystallographica: Section D (Wiley-Blackwell)
Publication Type :
Academic Journal
Accession number :
20870355
Full Text :
https://doi.org/10.1107/S0907444906010687