2hor
From Proteopedia
(New page: 200px<br /><applet load="2hor" size="450" color="white" frame="true" align="right" spinBox="true" caption="2hor, resolution 1.600Å" /> '''Crystal structure o...) |
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| - | [[Image:2hor.jpg|left|200px]]<br /><applet load="2hor" size=" | + | [[Image:2hor.jpg|left|200px]]<br /><applet load="2hor" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2hor, resolution 1.600Å" /> | caption="2hor, resolution 1.600Å" /> | ||
'''Crystal structure of alliinase from garlic- apo form'''<br /> | '''Crystal structure of alliinase from garlic- apo form'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Alliinase (alliin lyase EC 4.4.1.4), a PLP-dependent alpha, beta-eliminating lyase, constitutes one of the major protein components of | + | Alliinase (alliin lyase EC 4.4.1.4), a PLP-dependent alpha, beta-eliminating lyase, constitutes one of the major protein components of garlic (Alliium sativum L.) bulbs. The enzyme is a homodimeric glycoprotein and catalyzes the conversion of a specific non-protein sulfur-containing amino acid alliin ((+S)-allyl-L-cysteine sulfoxide) to allicin (diallyl thiosulfinate, the well known biologically active component of freshly crushed garlic), pyruvate and ammonia. The enzyme was crystallized in the presence of (+S)-allyl-L-cysteine, forming dendrite-like monoclinic crystals. In addition, intentionally produced apo-enzyme was crystallized in tetragonal form. These structures of alliinase with associated glycans were resolved to 1.4 A and 1.61 A by molecular replacement. Branched hexasaccharide chains N-linked to Asn146 and trisaccharide chains N-linked to Asn328 are seen. The structure of hexasaccharide was found similar to "short chain complex vacuole type" oligosaccharide most commonly seen in plant glycoproteins. An unexpected state of the enzyme active site has been observed in the present structure. The electron density in the region of the cofactor made it possible to identify the cofactor moiety as aminoacrylate intermediate covalently bound to the PLP cofactor. It was found in the present structure to be stabilized by large number of interactions with surrounding protein residues. Moreover, the existence of the expected internal aldimine bond between the epsilon-amino group of Lys251 and the aldehyde of the PLP is ruled out on the basis of a distinct separation of electron density of Lys251. The structure of the active site cavity in the apo-form is nearly identical to that seen in the holo-form, with two sulfate ions, an acetate and several water molecules from crystallization conditions that replace and mimic the PLP cofactor. |
==About this Structure== | ==About this Structure== | ||
| - | 2HOR is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Allium_sativum Allium sativum] with NAG, SO4, ACT, CL and NO3 as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alliin_lyase Alliin lyase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.4.1.4 4.4.1.4] Full crystallographic information is available from [http:// | + | 2HOR is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Allium_sativum Allium sativum] with <scene name='pdbligand=NAG:'>NAG</scene>, <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=ACT:'>ACT</scene>, <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=NO3:'>NO3</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alliin_lyase Alliin lyase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.4.1.4 4.4.1.4] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2HOR OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Mirelman, D.]] | [[Category: Mirelman, D.]] | ||
[[Category: Rabinkov, A.]] | [[Category: Rabinkov, A.]] | ||
| - | [[Category: Shimon, L | + | [[Category: Shimon, L J.W]] |
[[Category: Wilcheck, M.]] | [[Category: Wilcheck, M.]] | ||
[[Category: ACT]] | [[Category: ACT]] | ||
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[[Category: alliinase; garlic; allium sativum; glycosylation; plant enzyme; pyridoxal-5'-phosphate; aminoacrylate; apo form;]] | [[Category: alliinase; garlic; allium sativum; glycosylation; plant enzyme; pyridoxal-5'-phosphate; aminoacrylate; apo form;]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:44:03 2008'' |
Revision as of 15:44, 21 February 2008
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Crystal structure of alliinase from garlic- apo form
Overview
Alliinase (alliin lyase EC 4.4.1.4), a PLP-dependent alpha, beta-eliminating lyase, constitutes one of the major protein components of garlic (Alliium sativum L.) bulbs. The enzyme is a homodimeric glycoprotein and catalyzes the conversion of a specific non-protein sulfur-containing amino acid alliin ((+S)-allyl-L-cysteine sulfoxide) to allicin (diallyl thiosulfinate, the well known biologically active component of freshly crushed garlic), pyruvate and ammonia. The enzyme was crystallized in the presence of (+S)-allyl-L-cysteine, forming dendrite-like monoclinic crystals. In addition, intentionally produced apo-enzyme was crystallized in tetragonal form. These structures of alliinase with associated glycans were resolved to 1.4 A and 1.61 A by molecular replacement. Branched hexasaccharide chains N-linked to Asn146 and trisaccharide chains N-linked to Asn328 are seen. The structure of hexasaccharide was found similar to "short chain complex vacuole type" oligosaccharide most commonly seen in plant glycoproteins. An unexpected state of the enzyme active site has been observed in the present structure. The electron density in the region of the cofactor made it possible to identify the cofactor moiety as aminoacrylate intermediate covalently bound to the PLP cofactor. It was found in the present structure to be stabilized by large number of interactions with surrounding protein residues. Moreover, the existence of the expected internal aldimine bond between the epsilon-amino group of Lys251 and the aldehyde of the PLP is ruled out on the basis of a distinct separation of electron density of Lys251. The structure of the active site cavity in the apo-form is nearly identical to that seen in the holo-form, with two sulfate ions, an acetate and several water molecules from crystallization conditions that replace and mimic the PLP cofactor.
About this Structure
2HOR is a Single protein structure of sequence from Allium sativum with , , , and as ligands. Active as Alliin lyase, with EC number 4.4.1.4 Full crystallographic information is available from OCA.
Reference
Two structures of alliinase from Alliium sativum L.: apo form and ternary complex with aminoacrylate reaction intermediate covalently bound to the PLP cofactor., Shimon LJ, Rabinkov A, Shin I, Miron T, Mirelman D, Wilchek M, Frolow F, J Mol Biol. 2007 Feb 16;366(2):611-25. Epub 2006 Nov 14. PMID:17174334
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