2atm: Difference between revisions

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[[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size="350" color="white" frame="true" align="right" spinBox="true"

caption="2atm, resolution 2.000Å" />

'''Crystal structure of the recombinant allergen Ves v 2'''<br />

==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.

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 [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://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=2ATM OCA].

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==

Line 14:

[[Category: Single protein]]

[[Category: Vespula vulgaris]]

[[Category: Coen, J.J.F.]]

[[Category: Coen, J J.F.]]

[[Category: Crickmore, N.]]

[[Category: Gajhede, M.]]

[[Category: Kastrup, J.S.]]

[[Category: Kastrup, J S.]]

[[Category: King, T.P.]]

[[Category: King, T P.]]

[[Category: Monsalve, R.]]

[[Category: Seppala, U.]]

[[Category: Skov, L.K.]]

[[Category: Skov, L K.]]

[[Category: Spangfort, M.D.]]

[[Category: Spangfort, M D.]]

[[Category: MES]]

[[Category: SO4]]

[[Category: beta-alpha-barrels]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 08:22:~~05 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:30:57 2008''

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@@ Line 1: / Line 1: @@
-[[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:2atm.gif|left|200px]]<br /><applet load="2atm" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="2atm, resolution 2.000&Aring;" />
 '''Crystal structure of the recombinant allergen Ves v 2'''<br />
 ==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.
+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==
-ATM 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://ispc.weizmann.ac.il/oca-bin/ocashort?id=2ATM OCA].
+ATM 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==
@@ Line 14: / Line 14: @@
 [[Category: Single protein]]
 [[Category: Vespula vulgaris]]
-[[Category: Coen, J.J.F.]]
+[[Category: Coen, J J.F.]]
 [[Category: Crickmore, N.]]
 [[Category: Gajhede, M.]]
-[[Category: Kastrup, J.S.]]
+[[Category: Kastrup, J S.]]
-[[Category: King, T.P.]]
+[[Category: King, T P.]]
 [[Category: Monsalve, R.]]
 [[Category: Seppala, U.]]
-[[Category: Skov, L.K.]]
+[[Category: Skov, L K.]]
-[[Category: Spangfort, M.D.]]
+[[Category: Spangfort, M D.]]
 [[Category: MES]]
 [[Category: SO4]]
 [[Category: beta-alpha-barrels]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 08:22:05 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:30:57 2008''