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

Skov, L K, Seppala, U, Coen, J J F, Crickmore, N, King, T P, Monsalve, R, Kastrup, J S, Spangfort, M D and Gajhede, M (2006) Structure of recombinant Ves v 2 at 2.0 A resolution: structural analysis of an allergenic hyaluronidase from wasp venom. Acta Crystallographica Section D: Biological Crystallography, 62 (6). pp. 595-604. ISSN 0907-4449

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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 (A) over circle resolution and reveals a central ( beta/alpha)(7) 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.

Item Type: Article
Additional Information: Co-author. Coen was my post-doc. Result of EU collaborative grant.
Schools and Departments: School of Life Sciences > Biochemistry
Depositing User: Neil Crickmore
Date Deposited: 06 Feb 2012 18:30
Last Modified: 07 Jun 2012 09:00
URI: http://sro.sussex.ac.uk/id/eprint/16842
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