1zxi
From Proteopedia
(New page: 200px<br /><applet load="1zxi" size="450" color="white" frame="true" align="right" spinBox="true" caption="1zxi, resolution 1.7Å" /> '''Reconstituted CO dehy...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:1zxi.gif|left|200px]]<br /><applet load="1zxi" size=" | + | [[Image:1zxi.gif|left|200px]]<br /><applet load="1zxi" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1zxi, resolution 1.7Å" /> | caption="1zxi, resolution 1.7Å" /> | ||
'''Reconstituted CO dehydrogenase from Oligotropha carboxidovorans'''<br /> | '''Reconstituted CO dehydrogenase from Oligotropha carboxidovorans'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Carbon monoxide dehydrogenase from the bacterium Oligotropha | + | Carbon monoxide dehydrogenase from the bacterium Oligotropha carboxidovorans catalyzes the oxidation of CO to CO(2) at a unique [CuSMoO(2)] cluster. In the bacteria the cluster is assembled post-translational. The integration of S, and particularly of Cu, is rate limiting in vivo, which leads to CO dehydrogenase preparations containing the mature and fully functional enzyme along with forms of the enzyme deficient in one or both of these elements. The active sites of mature and immature forms of CO dehydrogenase were converted into a [MoO(3)] centre by treatment with potassium cyanide. We have established a method, which rescues 50% of the CO dehydrogenase activity by in vitro reconstitution of the active site through the supply of sulphide first and subsequently of Cu(I) under reducing conditions. Immature forms of CO dehydrogenase isolated from the bacterium, which were deficient in S and/or Cu at the active site, were similarly activated. X-ray crystallography and electron paramagnetic resonance spectroscopy indicated that the [CuSMoO(2)] cluster was properly reconstructed. However, reconstituted CO dehydrogenase contains mature along with immature forms. The chemical reactions of the reconstitution of CO dehydrogenase are summarized in a model, which assumes resulphuration of the Mo-ion at both equatorial positions at a 1:1 molar ratio. One equatorial Mo-S group reacts with Cu(I) in a productive fashion yielding a mature, functional [CuSMoO(2)] cluster. The other Mo-S group reacts with Cu(I), then Cu(2)S is released and an oxo group is introduced from water, yielding an inactive [MoO(3)] centre. |
==About this Structure== | ==About this Structure== | ||
| - | 1ZXI is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Oligotropha_carboxidovorans Oligotropha carboxidovorans] with CU, PO4, FES, CUM, MCN and FAD as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Carbon-monoxide_dehydrogenase_(acceptor) Carbon-monoxide dehydrogenase (acceptor)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.99.2 1.2.99.2] Full crystallographic information is available from [http:// | + | 1ZXI is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Oligotropha_carboxidovorans Oligotropha carboxidovorans] with <scene name='pdbligand=CU:'>CU</scene>, <scene name='pdbligand=PO4:'>PO4</scene>, <scene name='pdbligand=FES:'>FES</scene>, <scene name='pdbligand=CUM:'>CUM</scene>, <scene name='pdbligand=MCN:'>MCN</scene> and <scene name='pdbligand=FAD:'>FAD</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Carbon-monoxide_dehydrogenase_(acceptor) Carbon-monoxide dehydrogenase (acceptor)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.99.2 1.2.99.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1ZXI OCA]. |
==Reference== | ==Reference== | ||
| Line 27: | Line 27: | ||
[[Category: molybdoprotein]] | [[Category: molybdoprotein]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:20:03 2008'' |
Revision as of 14:20, 21 February 2008
|
Reconstituted CO dehydrogenase from Oligotropha carboxidovorans
Overview
Carbon monoxide dehydrogenase from the bacterium Oligotropha carboxidovorans catalyzes the oxidation of CO to CO(2) at a unique [CuSMoO(2)] cluster. In the bacteria the cluster is assembled post-translational. The integration of S, and particularly of Cu, is rate limiting in vivo, which leads to CO dehydrogenase preparations containing the mature and fully functional enzyme along with forms of the enzyme deficient in one or both of these elements. The active sites of mature and immature forms of CO dehydrogenase were converted into a [MoO(3)] centre by treatment with potassium cyanide. We have established a method, which rescues 50% of the CO dehydrogenase activity by in vitro reconstitution of the active site through the supply of sulphide first and subsequently of Cu(I) under reducing conditions. Immature forms of CO dehydrogenase isolated from the bacterium, which were deficient in S and/or Cu at the active site, were similarly activated. X-ray crystallography and electron paramagnetic resonance spectroscopy indicated that the [CuSMoO(2)] cluster was properly reconstructed. However, reconstituted CO dehydrogenase contains mature along with immature forms. The chemical reactions of the reconstitution of CO dehydrogenase are summarized in a model, which assumes resulphuration of the Mo-ion at both equatorial positions at a 1:1 molar ratio. One equatorial Mo-S group reacts with Cu(I) in a productive fashion yielding a mature, functional [CuSMoO(2)] cluster. The other Mo-S group reacts with Cu(I), then Cu(2)S is released and an oxo group is introduced from water, yielding an inactive [MoO(3)] centre.
About this Structure
1ZXI is a Protein complex structure of sequences from Oligotropha carboxidovorans with , , , , and as ligands. Active as Carbon-monoxide dehydrogenase (acceptor), with EC number 1.2.99.2 Full crystallographic information is available from OCA.
Reference
Structural and functional reconstruction in situ of the [CuSMoO2] active site of carbon monoxide dehydrogenase from the carbon monoxide oxidizing eubacterium Oligotropha carboxidovorans., Resch M, Dobbek H, Meyer O, J Biol Inorg Chem. 2005 Aug;10(5):518-28. Epub 2005 Sep 23. PMID:16091936
Page seeded by OCA on Thu Feb 21 16:20:03 2008
Categories: Carbon-monoxide dehydrogenase (acceptor) | Oligotropha carboxidovorans | Protein complex | Dobbek, H. | Meyer, O. | Resch, M. | CU | CUM | FAD | FES | MCN | PO4 | Codh | Molybdenum | Molybdoprotein
