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 journalJournal articleResearchpeer-review

Harvard

Skov, LK, Seppälä, U, Coen, JJF, Crickmore, N, King, TP, Monsalve, R, Kastrup, JS, Spangfort, MD & Gajhede, M 2006, 'Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom', Acta Crystallographica. Section D: Biological Crystallography, vol. 62, no. Pt 6, pp. 595-604. https://doi.org/10.1107/S0907444906010687

APA

Skov, L. K., Seppälä, U., Coen, J. J. F., Crickmore, N., King, T. P., Monsalve, R., ... Gajhede, M. (2006). Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom. Acta Crystallographica. Section D: Biological Crystallography, 62(Pt 6), 595-604. https://doi.org/10.1107/S0907444906010687

Vancouver

Skov LK, Seppälä U, Coen JJF, Crickmore N, King TP, Monsalve R et al. Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom. Acta Crystallographica. Section D: Biological Crystallography. 2006;62(Pt 6):595-604. https://doi.org/10.1107/S0907444906010687

Author

Skov, Lars K ; Seppälä, Ulla ; Coen, Jeremy J F ; Crickmore, Neil ; King, Te P ; Monsalve, Rafael ; Kastrup, Jette S ; Spangfort, Michael D ; Gajhede, Michael. / Structure of recombinant Ves v 2 at 2.0 Angstrom resolution : structural analysis of an allergenic hyaluronidase from wasp venom. In: Acta Crystallographica. Section D: Biological Crystallography. 2006 ; Vol. 62, No. Pt 6. pp. 595-604.

Bibtex

@article{e1a94cfc0c6b4e878bfb712bbaf51fd3,
title = "Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom",
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 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.",
keywords = "Amino Acid Sequence, Binding Sites, Crystallography, X-Ray, Glycosylation, Hyaluronoglucosaminidase, Models, Molecular, Molecular Sequence Data, Protein Folding, Recombinant Proteins, Sequence Alignment, Sequence Analysis, Protein, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Wasp Venoms",
author = "Skov, {Lars K} and Ulla Sepp{\"a}l{\"a} and Coen, {Jeremy J F} and Neil Crickmore and King, {Te P} and Rafael Monsalve and Kastrup, {Jette S} and Spangfort, {Michael D} and Michael Gajhede",
year = "2006",
doi = "10.1107/S0907444906010687",
language = "English",
volume = "62",
pages = "595--604",
journal = "Acta Crystallographica Section D: Structural Biology",
issn = "2059-7983",
publisher = "International Union of Crystallography",
number = "Pt 6",

}

RIS

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