1ks9
From Proteopedia
(New page: 200px<br /><applet load="1ks9" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ks9, resolution 1.70Å" /> '''Ketopantoate Reducta...) |
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| - | [[Image:1ks9.jpg|left|200px]]<br /><applet load="1ks9" size=" | + | [[Image:1ks9.jpg|left|200px]]<br /><applet load="1ks9" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1ks9, resolution 1.70Å" /> | caption="1ks9, resolution 1.70Å" /> | ||
'''Ketopantoate Reductase from Escherichia coli'''<br /> | '''Ketopantoate Reductase from Escherichia coli'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Ketopantoate reductase (KPR, EC 1.1.1.169) catalyzes the NADPH-dependent | + | Ketopantoate reductase (KPR, EC 1.1.1.169) catalyzes the NADPH-dependent reduction of ketopantoate to pantoate on the pantothenate (vitamin B(5)) biosynthetic pathway. The Escherichia coli panE gene encoding KPR was cloned and expressed at high levels as the native and selenomethionine-substituted (SeMet) proteins. Both native and SeMet recombinant proteins were purified by three chromatographic steps, to yield pure proteins. The wild-type enzyme was found to have a K(M)(NADPH) of 20 microM, a K(M)(ketopantoate) of 60 microM, and a k(cat) of 40 s(-1). Regular prismatic KPR crystals were prepared using the hanging drop technique. They belonged to the tetragonal space group P4(2)2(1)2, with cell parameters: a = b = 103.7 A and c = 55.7 A, accommodating one enzyme molecule per asymmetric unit. The structure of KPR was determined by the multiwavelength anomalous dispersion method using the SeMet protein, for which data were collected to 2.3 A resolution. The native data were collected to 1.7 A resolution and used to refine the final structure. The secondary structure comprises 12 alpha-helices, three 3(10)-helices, and 11 beta-strands. The enzyme is monomeric and has two domains separated by a cleft. The N-terminal domain has an alphabeta-fold of the Rossmann type. The C-terminal domain (residues 170-291) is composed of eight alpha-helices. KPR is shown to be a member of the 6-phosphogluconate dehydrogenase C-terminal domain-like superfamily. A model for the ternary enzyme-NADPH-ketopantoate ternary complex provides a rationale for kinetic data reported for specific site-directed mutants. |
==About this Structure== | ==About this Structure== | ||
| - | 1KS9 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Active as [http://en.wikipedia.org/wiki/2-dehydropantoate_2-reductase 2-dehydropantoate 2-reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.169 1.1.1.169] Full crystallographic information is available from [http:// | + | 1KS9 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Active as [http://en.wikipedia.org/wiki/2-dehydropantoate_2-reductase 2-dehydropantoate 2-reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.169 1.1.1.169] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1KS9 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Abell, C.]] | [[Category: Abell, C.]] | ||
| - | [[Category: Ashurst, J | + | [[Category: Ashurst, J A.]] |
| - | [[Category: Blundell, T | + | [[Category: Blundell, T L.]] |
| - | [[Category: Delft, F | + | [[Category: Delft, F von.]] |
[[Category: Inoue, T.]] | [[Category: Inoue, T.]] | ||
[[Category: Matak-Vinkovic, D.]] | [[Category: Matak-Vinkovic, D.]] | ||
| - | [[Category: Miguel, R | + | [[Category: Miguel, R N.]] |
| - | [[Category: Saldanha, S | + | [[Category: Saldanha, S A.]] |
| - | [[Category: Smith, A | + | [[Category: Smith, A G.]] |
[[Category: Vinkovic, M.]] | [[Category: Vinkovic, M.]] | ||
[[Category: apba]] | [[Category: apba]] | ||
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[[Category: rossman fold]] | [[Category: rossman fold]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:37:23 2008'' |
Revision as of 11:37, 21 February 2008
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Ketopantoate Reductase from Escherichia coli
Overview
Ketopantoate reductase (KPR, EC 1.1.1.169) catalyzes the NADPH-dependent reduction of ketopantoate to pantoate on the pantothenate (vitamin B(5)) biosynthetic pathway. The Escherichia coli panE gene encoding KPR was cloned and expressed at high levels as the native and selenomethionine-substituted (SeMet) proteins. Both native and SeMet recombinant proteins were purified by three chromatographic steps, to yield pure proteins. The wild-type enzyme was found to have a K(M)(NADPH) of 20 microM, a K(M)(ketopantoate) of 60 microM, and a k(cat) of 40 s(-1). Regular prismatic KPR crystals were prepared using the hanging drop technique. They belonged to the tetragonal space group P4(2)2(1)2, with cell parameters: a = b = 103.7 A and c = 55.7 A, accommodating one enzyme molecule per asymmetric unit. The structure of KPR was determined by the multiwavelength anomalous dispersion method using the SeMet protein, for which data were collected to 2.3 A resolution. The native data were collected to 1.7 A resolution and used to refine the final structure. The secondary structure comprises 12 alpha-helices, three 3(10)-helices, and 11 beta-strands. The enzyme is monomeric and has two domains separated by a cleft. The N-terminal domain has an alphabeta-fold of the Rossmann type. The C-terminal domain (residues 170-291) is composed of eight alpha-helices. KPR is shown to be a member of the 6-phosphogluconate dehydrogenase C-terminal domain-like superfamily. A model for the ternary enzyme-NADPH-ketopantoate ternary complex provides a rationale for kinetic data reported for specific site-directed mutants.
About this Structure
1KS9 is a Single protein structure of sequence from Escherichia coli. Active as 2-dehydropantoate 2-reductase, with EC number 1.1.1.169 Full crystallographic information is available from OCA.
Reference
Crystal structure of Escherichia coli ketopantoate reductase at 1.7 A resolution and insight into the enzyme mechanism., Matak-Vinkovic D, Vinkovic M, Saldanha SA, Ashurst JL, von Delft F, Inoue T, Miguel RN, Smith AG, Blundell TL, Abell C, Biochemistry. 2001 Dec 4;40(48):14493-500. PMID:11724562
Page seeded by OCA on Thu Feb 21 13:37:23 2008
Categories: 2-dehydropantoate 2-reductase | Escherichia coli | Single protein | Abell, C. | Ashurst, J A. | Blundell, T L. | Delft, F von. | Inoue, T. | Matak-Vinkovic, D. | Miguel, R N. | Saldanha, S A. | Smith, A G. | Vinkovic, M. | Apba | Apo | Ketopantoate reductase | Monomer | Pane | Rossman fold
