2atm
From Proteopedia
(New page: 200px<br /><applet load="2atm" size="450" color="white" frame="true" align="right" spinBox="true" caption="2atm, resolution 2.000Å" /> '''Crystal structure o...) |
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| - | [[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size=" | + | [[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2atm, resolution 2.000Å" /> | caption="2atm, resolution 2.000Å" /> | ||
'''Crystal structure of the recombinant allergen Ves v 2'''<br /> | '''Crystal structure of the recombinant allergen Ves v 2'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Wasp venom from Vespula vulgaris contains three major allergens: Ves v 1, Ves v 2 and Ves v 5. Here, the cloning, expression, biochemical | + | 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. |
==About this Structure== | ==About this Structure== | ||
| - | 2ATM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Vespula_vulgaris Vespula vulgaris] with SO4 and MES as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Hyalurononglucosaminidase Hyalurononglucosaminidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.35 3.2.1.35] Full crystallographic information is available from [http:// | + | 2ATM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Vespula_vulgaris Vespula vulgaris] with <scene name='pdbligand=SO4:'>SO4</scene> and <scene name='pdbligand=MES:'>MES</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Hyalurononglucosaminidase Hyalurononglucosaminidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.35 3.2.1.35] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2ATM OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Vespula vulgaris]] | [[Category: Vespula vulgaris]] | ||
| - | [[Category: Coen, J | + | [[Category: Coen, J J.F.]] |
[[Category: Crickmore, N.]] | [[Category: Crickmore, N.]] | ||
[[Category: Gajhede, M.]] | [[Category: Gajhede, M.]] | ||
| - | [[Category: Kastrup, J | + | [[Category: Kastrup, J S.]] |
| - | [[Category: King, T | + | [[Category: King, T P.]] |
[[Category: Monsalve, R.]] | [[Category: Monsalve, R.]] | ||
[[Category: Seppala, U.]] | [[Category: Seppala, U.]] | ||
| - | [[Category: Skov, L | + | [[Category: Skov, L K.]] |
| - | [[Category: Spangfort, M | + | [[Category: Spangfort, M D.]] |
[[Category: MES]] | [[Category: MES]] | ||
[[Category: SO4]] | [[Category: SO4]] | ||
[[Category: beta-alpha-barrels]] | [[Category: beta-alpha-barrels]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:30:57 2008'' |
Revision as of 14:31, 21 February 2008
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Crystal structure of the recombinant allergen Ves v 2
Overview
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.
About this Structure
2ATM is a Single protein structure of sequence from Vespula vulgaris with and as ligands. Active as Hyalurononglucosaminidase, with EC number 3.2.1.35 Full crystallographic information is available from OCA.
Reference
Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom., Skov LK, Seppala U, Coen JJ, Crickmore N, King TP, Monsalve R, Kastrup JS, Spangfort MD, Gajhede M, Acta Crystallogr D Biol Crystallogr. 2006 Jun;62(Pt 6):595-604. Epub 2006, May 12. PMID:16699186
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