Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom
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Structure of recombinant Ves v 2 at 2.0 Angstrom resolution : structural analysis of an allergenic hyaluronidase from wasp venom. / Skov, Lars K; Seppälä, Ulla; Coen, Jeremy J F; Crickmore, Neil; King, Te P; Monsalve, Rafael; Kastrup, Jette S; Spangfort, Michael D; Gajhede, Michael.
In: Acta Crystallographica. Section D: Biological Crystallography, Vol. 62, No. Pt 6, 2006, p. 595-604.Research output: Contribution to journal › Journal article › peer-review
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TY - JOUR
T1 - Structure of recombinant Ves v 2 at 2.0 Angstrom resolution
T2 - structural analysis of an allergenic hyaluronidase from wasp venom
AU - Skov, Lars K
AU - Seppälä, Ulla
AU - Coen, Jeremy J F
AU - Crickmore, Neil
AU - King, Te P
AU - Monsalve, Rafael
AU - Kastrup, Jette S
AU - Spangfort, Michael D
AU - Gajhede, Michael
PY - 2006
Y1 - 2006
N2 - 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 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.
AB - 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 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.
KW - Amino Acid Sequence
KW - Binding Sites
KW - Crystallography, X-Ray
KW - Glycosylation
KW - Hyaluronoglucosaminidase
KW - Models, Molecular
KW - Molecular Sequence Data
KW - Protein Folding
KW - Recombinant Proteins
KW - Sequence Alignment
KW - Sequence Analysis, Protein
KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
KW - Wasp Venoms
U2 - 10.1107/S0907444906010687
DO - 10.1107/S0907444906010687
M3 - Journal article
C2 - 16699186
VL - 62
SP - 595
EP - 604
JO - Acta Crystallographica Section D: Structural Biology
JF - Acta Crystallographica Section D: Structural Biology
SN - 2059-7983
IS - Pt 6
ER -
ID: 40766626