1yrc
From Proteopedia
(New page: 200px<br /><applet load="1yrc" size="450" color="white" frame="true" align="right" spinBox="true" caption="1yrc, resolution 1.4Å" /> '''X-ray Crystal Structu...) |
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| - | [[Image:1yrc.gif|left|200px]]<br /><applet load="1yrc" size=" | + | [[Image:1yrc.gif|left|200px]]<br /><applet load="1yrc" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1yrc, resolution 1.4Å" /> | caption="1yrc, resolution 1.4Å" /> | ||
'''X-ray Crystal Structure of hydrogenated Cytochrome P450cam'''<br /> | '''X-ray Crystal Structure of hydrogenated Cytochrome P450cam'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Neutron protein crystallography allows H-atom positions to be located in | + | Neutron protein crystallography allows H-atom positions to be located in biological structures at the relatively modest resolution of 1.5-2.0 A. A difficulty of this technique arises from the incoherent scattering from hydrogen, which considerably reduces the signal-to-noise ratio of the data. This can be overcome by preparing fully deuterated samples. Efficient protocols for routine and low-cost production of in vivo deuterium-enriched proteins have been developed. Here, the overexpression and crystallization of highly (>99%) deuterium-enriched cytochrome P450cam for neutron analysis is reported. Cytochrome P450cam from Pseudomonas putida catalyses the hydroxylation of camphor from haem-bound molecular O(2) via a mechanism that is thought to involve a proton-shuttle pathway to the active site. Since H atoms cannot be visualized in available X-ray structures, neutron diffraction is being used to determine the protonation states and water structure at the active site of the enzyme. Analysis of both hydrogenated and perdeuterated P450cam showed no significant changes between the X-ray structures determined at 1.4 and 1.7 A, respectively. This work demonstrates that the fully deuterated protein is highly isomorphous with the native (hydrogenated) protein and is appropriate for neutron protein crystallographic analysis. |
==About this Structure== | ==About this Structure== | ||
| - | 1YRC is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida] with K, HEM and CAM as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Camphor_5-monooxygenase Camphor 5-monooxygenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.15.1 1.14.15.1] Full crystallographic information is available from [http:// | + | 1YRC is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida] with <scene name='pdbligand=K:'>K</scene>, <scene name='pdbligand=HEM:'>HEM</scene> and <scene name='pdbligand=CAM:'>CAM</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Camphor_5-monooxygenase Camphor 5-monooxygenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.15.1 1.14.15.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1YRC OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Pseudomonas putida]] | [[Category: Pseudomonas putida]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Dauvergne, M | + | [[Category: Dauvergne, M T.]] |
[[Category: Meilleur, F.]] | [[Category: Meilleur, F.]] | ||
| - | [[Category: Myles, D | + | [[Category: Myles, D A.A.]] |
[[Category: Schlichting, I.]] | [[Category: Schlichting, I.]] | ||
[[Category: CAM]] | [[Category: CAM]] | ||
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[[Category: oxidoreductase]] | [[Category: oxidoreductase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:08:17 2008'' |
Revision as of 14:08, 21 February 2008
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X-ray Crystal Structure of hydrogenated Cytochrome P450cam
Overview
Neutron protein crystallography allows H-atom positions to be located in biological structures at the relatively modest resolution of 1.5-2.0 A. A difficulty of this technique arises from the incoherent scattering from hydrogen, which considerably reduces the signal-to-noise ratio of the data. This can be overcome by preparing fully deuterated samples. Efficient protocols for routine and low-cost production of in vivo deuterium-enriched proteins have been developed. Here, the overexpression and crystallization of highly (>99%) deuterium-enriched cytochrome P450cam for neutron analysis is reported. Cytochrome P450cam from Pseudomonas putida catalyses the hydroxylation of camphor from haem-bound molecular O(2) via a mechanism that is thought to involve a proton-shuttle pathway to the active site. Since H atoms cannot be visualized in available X-ray structures, neutron diffraction is being used to determine the protonation states and water structure at the active site of the enzyme. Analysis of both hydrogenated and perdeuterated P450cam showed no significant changes between the X-ray structures determined at 1.4 and 1.7 A, respectively. This work demonstrates that the fully deuterated protein is highly isomorphous with the native (hydrogenated) protein and is appropriate for neutron protein crystallographic analysis.
About this Structure
1YRC is a Single protein structure of sequence from Pseudomonas putida with , and as ligands. Active as Camphor 5-monooxygenase, with EC number 1.14.15.1 Full crystallographic information is available from OCA.
Reference
Production and X-ray crystallographic analysis of fully deuterated cytochrome P450cam., Meilleur F, Dauvergne MT, Schlichting I, Myles DA, Acta Crystallogr D Biol Crystallogr. 2005 May;61(Pt 5):539-44. Epub 2005, Apr 20. PMID:15858263
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