2f17

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(New page: 200px<br /><applet load="2f17" size="450" color="white" frame="true" align="right" spinBox="true" caption="2f17, resolution 2.5&Aring;" /> '''Mouse Thiamin Pyropho...)
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[[Image:2f17.gif|left|200px]]<br /><applet load="2f17" size="350" color="white" frame="true" align="right" spinBox="true"
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caption="2f17, resolution 2.5&Aring;" />
'''Mouse Thiamin Pyrophosphokinase in a Ternary Complex with Pyrithiamin Pyrophosphate and AMP at 2.5 angstrom'''<br />
'''Mouse Thiamin Pyrophosphokinase in a Ternary Complex with Pyrithiamin Pyrophosphate and AMP at 2.5 angstrom'''<br />
==Overview==
==Overview==
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Thiamine pyrophosphokinase transfers a pyrophosphate group from a, nucleoside triphosphate, such as ATP, to the hydroxyl group of thiamine to, produce thiamine pyrophosphate. Deficiencies in thiamine can result in the, development of the neurological disorder Wernicke-Korsakoff Syndrome as, well as the potentially fatal cardiovascular disease wet beriberi., Pyrithiamine is an inhibitor of thiamine metabolism that induces, neurological symptoms similar to that of Wernicke-Korsakoff Syndrome in, animals. However, the mechanism by which pyrithiamine interferes with, cellular thiamine phosphoester homeostasis is not entirely clear. We used, kinetic assays coupled with mass spectrometry of the reaction products and, x-ray crystallography of an equilibrium reaction mixture of thiamine, pyrophosphokinase, pyrithiamine, and Mg2+/ATP to elucidate the mechanism, by which pyrithiamine inhibits the enzymatic production of thiamine, pyrophosphate. Three lines of evidence support the ability of thiamine, pyrophosphokinase to form pyrithiamine pyrophosphate. First, a coupled, enzyme assay clearly demonstrated the ability of thiamine, pyrophosphokinase to produce AMP when pyrithiamine was used as substrate., Second, an analysis of the reaction mixture by mass spectrometry directly, identified pyrithiamine pyrophosphate in the reaction mixture. Last, the, structure of thiamine pyrophosphokinase crystallized from an equilibrium, substrate/product mixture shows clear electron density for pyrithiamine, pyrophosphate bound in the enzyme active site. This structure also, provides the first clear picture of the binding pocket for the nucleoside, triphosphate and permits the first detailed understanding of the catalytic, requirements for catalysis in this enzyme.
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Thiamine pyrophosphokinase transfers a pyrophosphate group from a nucleoside triphosphate, such as ATP, to the hydroxyl group of thiamine to produce thiamine pyrophosphate. Deficiencies in thiamine can result in the development of the neurological disorder Wernicke-Korsakoff Syndrome as well as the potentially fatal cardiovascular disease wet beriberi. Pyrithiamine is an inhibitor of thiamine metabolism that induces neurological symptoms similar to that of Wernicke-Korsakoff Syndrome in animals. However, the mechanism by which pyrithiamine interferes with cellular thiamine phosphoester homeostasis is not entirely clear. We used kinetic assays coupled with mass spectrometry of the reaction products and x-ray crystallography of an equilibrium reaction mixture of thiamine pyrophosphokinase, pyrithiamine, and Mg2+/ATP to elucidate the mechanism by which pyrithiamine inhibits the enzymatic production of thiamine pyrophosphate. Three lines of evidence support the ability of thiamine pyrophosphokinase to form pyrithiamine pyrophosphate. First, a coupled enzyme assay clearly demonstrated the ability of thiamine pyrophosphokinase to produce AMP when pyrithiamine was used as substrate. Second, an analysis of the reaction mixture by mass spectrometry directly identified pyrithiamine pyrophosphate in the reaction mixture. Last, the structure of thiamine pyrophosphokinase crystallized from an equilibrium substrate/product mixture shows clear electron density for pyrithiamine pyrophosphate bound in the enzyme active site. This structure also provides the first clear picture of the binding pocket for the nucleoside triphosphate and permits the first detailed understanding of the catalytic requirements for catalysis in this enzyme.
==About this Structure==
==About this Structure==
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2F17 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus] with MG, SO4, EPE, PYI and AMP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Thiamine_diphosphokinase Thiamine diphosphokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.6.2 2.7.6.2] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2F17 OCA].
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2F17 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus] with <scene name='pdbligand=MG:'>MG</scene>, <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=EPE:'>EPE</scene>, <scene name='pdbligand=PYI:'>PYI</scene> and <scene name='pdbligand=AMP:'>AMP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Thiamine_diphosphokinase Thiamine diphosphokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.6.2 2.7.6.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2F17 OCA].
==Reference==
==Reference==
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[[Category: Single protein]]
[[Category: Single protein]]
[[Category: Thiamine diphosphokinase]]
[[Category: Thiamine diphosphokinase]]
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[[Category: Hurley, T.D.]]
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[[Category: Hurley, T D.]]
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[[Category: Liu, J.Y.]]
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[[Category: Liu, J Y.]]
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[[Category: Timm, D.E.]]
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[[Category: Timm, D E.]]
[[Category: AMP]]
[[Category: AMP]]
[[Category: EPE]]
[[Category: EPE]]
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[[Category: pyrophosphokinase]]
[[Category: pyrophosphokinase]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 10:17:36 2007''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:16:37 2008''

Revision as of 15:16, 21 February 2008

Image:2f17.gif

2f17, resolution 2.5Å

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Mouse Thiamin Pyrophosphokinase in a Ternary Complex with Pyrithiamin Pyrophosphate and AMP at 2.5 angstrom

Overview

Thiamine pyrophosphokinase transfers a pyrophosphate group from a nucleoside triphosphate, such as ATP, to the hydroxyl group of thiamine to produce thiamine pyrophosphate. Deficiencies in thiamine can result in the development of the neurological disorder Wernicke-Korsakoff Syndrome as well as the potentially fatal cardiovascular disease wet beriberi. Pyrithiamine is an inhibitor of thiamine metabolism that induces neurological symptoms similar to that of Wernicke-Korsakoff Syndrome in animals. However, the mechanism by which pyrithiamine interferes with cellular thiamine phosphoester homeostasis is not entirely clear. We used kinetic assays coupled with mass spectrometry of the reaction products and x-ray crystallography of an equilibrium reaction mixture of thiamine pyrophosphokinase, pyrithiamine, and Mg2+/ATP to elucidate the mechanism by which pyrithiamine inhibits the enzymatic production of thiamine pyrophosphate. Three lines of evidence support the ability of thiamine pyrophosphokinase to form pyrithiamine pyrophosphate. First, a coupled enzyme assay clearly demonstrated the ability of thiamine pyrophosphokinase to produce AMP when pyrithiamine was used as substrate. Second, an analysis of the reaction mixture by mass spectrometry directly identified pyrithiamine pyrophosphate in the reaction mixture. Last, the structure of thiamine pyrophosphokinase crystallized from an equilibrium substrate/product mixture shows clear electron density for pyrithiamine pyrophosphate bound in the enzyme active site. This structure also provides the first clear picture of the binding pocket for the nucleoside triphosphate and permits the first detailed understanding of the catalytic requirements for catalysis in this enzyme.

About this Structure

2F17 is a Single protein structure of sequence from Mus musculus with , , , and as ligands. Active as Thiamine diphosphokinase, with EC number 2.7.6.2 Full crystallographic information is available from OCA.

Reference

Pyrithiamine as a substrate for thiamine pyrophosphokinase., Liu JY, Timm DE, Hurley TD, J Biol Chem. 2006 Mar 10;281(10):6601-7. Epub 2005 Dec 19. PMID:16365036

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