2ez8
From Proteopedia
(New page: 200px<br /><applet load="2ez8" size="450" color="white" frame="true" align="right" spinBox="true" caption="2ez8, resolution 1.963Å" /> '''Pyruvate oxidase va...) |
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| - | [[Image:2ez8.gif|left|200px]]<br /><applet load="2ez8" size=" | + | [[Image:2ez8.gif|left|200px]]<br /><applet load="2ez8" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2ez8, resolution 1.963Å" /> | caption="2ez8, resolution 1.963Å" /> | ||
'''Pyruvate oxidase variant F479W in complex with reaction intermediate 2-lactyl-thiamin diphosphate'''<br /> | '''Pyruvate oxidase variant F479W in complex with reaction intermediate 2-lactyl-thiamin diphosphate'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Enzymes that use the cofactor thiamin diphosphate (ThDP, 1), the | + | Enzymes that use the cofactor thiamin diphosphate (ThDP, 1), the biologically active form of vitamin B(1), are involved in numerous metabolic pathways in all organisms. Although a theory of the cofactor's underlying reaction mechanism has been established over the last five decades, the three-dimensional structures of most major reaction intermediates of ThDP enzymes have remained elusive. Here, we report the X-ray structures of key intermediates in the oxidative decarboxylation of pyruvate, a central reaction in carbon metabolism catalyzed by the ThDP- and flavin-dependent enzyme pyruvate oxidase (POX)3 from Lactobacillus plantarum. The structures of 2-lactyl-ThDP (LThDP, 2) and its stable phosphonate analog, of 2-hydroxyethyl-ThDP (HEThDP, 3) enamine and of 2-acetyl-ThDP (AcThDP, 4; all shown bound to the enzyme's active site) provide profound insights into the chemical mechanisms and the stereochemical course of thiamin catalysis. These snapshots also suggest a mechanism for a phosphate-linked acyl transfer coupled to electron transfer in a radical reaction of pyruvate oxidase. |
==About this Structure== | ==About this Structure== | ||
| - | 2EZ8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactobacillus_plantarum Lactobacillus plantarum] with MG, NA, TDL, FAD and PYR as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Pyruvate_oxidase Pyruvate oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.3.3 1.2.3.3] Full crystallographic information is available from [http:// | + | 2EZ8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactobacillus_plantarum Lactobacillus plantarum] with <scene name='pdbligand=MG:'>MG</scene>, <scene name='pdbligand=NA:'>NA</scene>, <scene name='pdbligand=TDL:'>TDL</scene>, <scene name='pdbligand=FAD:'>FAD</scene> and <scene name='pdbligand=PYR:'>PYR</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Pyruvate_oxidase Pyruvate oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.3.3 1.2.3.3] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2EZ8 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: tpp enzyme]] | [[Category: tpp enzyme]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:16:02 2008'' |
Revision as of 15:16, 21 February 2008
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Pyruvate oxidase variant F479W in complex with reaction intermediate 2-lactyl-thiamin diphosphate
Overview
Enzymes that use the cofactor thiamin diphosphate (ThDP, 1), the biologically active form of vitamin B(1), are involved in numerous metabolic pathways in all organisms. Although a theory of the cofactor's underlying reaction mechanism has been established over the last five decades, the three-dimensional structures of most major reaction intermediates of ThDP enzymes have remained elusive. Here, we report the X-ray structures of key intermediates in the oxidative decarboxylation of pyruvate, a central reaction in carbon metabolism catalyzed by the ThDP- and flavin-dependent enzyme pyruvate oxidase (POX)3 from Lactobacillus plantarum. The structures of 2-lactyl-ThDP (LThDP, 2) and its stable phosphonate analog, of 2-hydroxyethyl-ThDP (HEThDP, 3) enamine and of 2-acetyl-ThDP (AcThDP, 4; all shown bound to the enzyme's active site) provide profound insights into the chemical mechanisms and the stereochemical course of thiamin catalysis. These snapshots also suggest a mechanism for a phosphate-linked acyl transfer coupled to electron transfer in a radical reaction of pyruvate oxidase.
About this Structure
2EZ8 is a Single protein structure of sequence from Lactobacillus plantarum with , , , and as ligands. Active as Pyruvate oxidase, with EC number 1.2.3.3 Full crystallographic information is available from OCA.
Reference
The catalytic cycle of a thiamin diphosphate enzyme examined by cryocrystallography., Wille G, Meyer D, Steinmetz A, Hinze E, Golbik R, Tittmann K, Nat Chem Biol. 2006 Jun;2(6):324-8. Epub 2006 May 7. PMID:16680160
Page seeded by OCA on Thu Feb 21 17:16:02 2008
Categories: Lactobacillus plantarum | Pyruvate oxidase | Single protein | Golbik, R. | Hinze, E. | Meyer, D. | Steinmetz, A. | Tittmann, K. | Wille, G. | FAD | MG | NA | PYR | TDL | Reaction intermediate | Tpp enzyme
