2nac
From Proteopedia
(New page: 200px<br /><applet load="2nac" size="450" color="white" frame="true" align="right" spinBox="true" caption="2nac, resolution 1.80Å" /> '''HIGH RESOLUTION STRU...) |
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| - | [[Image:2nac.jpg|left|200px]]<br /><applet load="2nac" size=" | + | [[Image:2nac.jpg|left|200px]]<br /><applet load="2nac" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2nac, resolution 1.80Å" /> | caption="2nac, resolution 1.80Å" /> | ||
'''HIGH RESOLUTION STRUCTURES OF HOLO AND APO FORMATE DEHYDROGENASE'''<br /> | '''HIGH RESOLUTION STRUCTURES OF HOLO AND APO FORMATE DEHYDROGENASE'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Three-dimensional crystal structures of holo (ternary complex | + | Three-dimensional crystal structures of holo (ternary complex enzyme-NAD-azide) and apo NAD-dependent dimeric formate dehydrogenase (FDH) from the methylotrophic bacterium Pseudomonas sp. 101 have been refined to R factors of 11.7% and 14.8% at 2.05 and 1.80 A resolution, respectively. The estimated root-mean-square error in atomic co-ordinates is 0.11 A for holo and 0.18 A for apo. X-ray data were collected from single crystals using an imaging plate scanner and synchrotron radiation. In both crystal forms there is a dimer in the asymmetric unit. Both structures show essentially 2-fold molecular symmetry. NAD binding causes movement of the catalytic domain and ordering of the C terminus, where a new helix appears. This completes formation of the enzyme active centre in holo FDH. NAD is bound in the cleft separating the domains and mainly interacts with residues from the co-enzyme binding domain. In apo FDH these residues are held in essentially the same conformation by water molecules occupying the NAD binding region. An azide molecule is located near the point of catalysis, the C4 atom of the nicotinamide moiety of NAD, and overlaps with the proposed formate binding site. There is an extensive channel running from the active site to the protein surface and this is supposed to be used by substrate to reach the active centre after NAD has already bound. The structure of the active site and a hypothetical catalytic mechanism are discussed. Sequence homology of FDH with other NAD-dependent formate dehydrogenases and some D-specific dehydrogenases is discussed on the basis of the FDH three-dimensional structure. |
==About this Structure== | ==About this Structure== | ||
| - | 2NAC is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_sp. Pseudomonas sp.] with SO4 as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Formate_dehydrogenase Formate dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.1.2 1.2.1.2] Full crystallographic information is available from [http:// | + | 2NAC is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_sp. Pseudomonas sp.] with <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Formate_dehydrogenase Formate dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.2.1.2 1.2.1.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2NAC OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Dauter, Z.]] | [[Category: Dauter, Z.]] | ||
| - | [[Category: Harutyunyan, E | + | [[Category: Harutyunyan, E H.]] |
| - | [[Category: Lamzin, V | + | [[Category: Lamzin, V S.]] |
| - | [[Category: Popov, V | + | [[Category: Popov, V O.]] |
| - | [[Category: Wilson, K | + | [[Category: Wilson, K S.]] |
[[Category: SO4]] | [[Category: SO4]] | ||
[[Category: nad+(a))]] | [[Category: nad+(a))]] | ||
[[Category: oxidoreductase(aldehyde(d)]] | [[Category: oxidoreductase(aldehyde(d)]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:08:06 2008'' |
Revision as of 16:08, 21 February 2008
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HIGH RESOLUTION STRUCTURES OF HOLO AND APO FORMATE DEHYDROGENASE
Overview
Three-dimensional crystal structures of holo (ternary complex enzyme-NAD-azide) and apo NAD-dependent dimeric formate dehydrogenase (FDH) from the methylotrophic bacterium Pseudomonas sp. 101 have been refined to R factors of 11.7% and 14.8% at 2.05 and 1.80 A resolution, respectively. The estimated root-mean-square error in atomic co-ordinates is 0.11 A for holo and 0.18 A for apo. X-ray data were collected from single crystals using an imaging plate scanner and synchrotron radiation. In both crystal forms there is a dimer in the asymmetric unit. Both structures show essentially 2-fold molecular symmetry. NAD binding causes movement of the catalytic domain and ordering of the C terminus, where a new helix appears. This completes formation of the enzyme active centre in holo FDH. NAD is bound in the cleft separating the domains and mainly interacts with residues from the co-enzyme binding domain. In apo FDH these residues are held in essentially the same conformation by water molecules occupying the NAD binding region. An azide molecule is located near the point of catalysis, the C4 atom of the nicotinamide moiety of NAD, and overlaps with the proposed formate binding site. There is an extensive channel running from the active site to the protein surface and this is supposed to be used by substrate to reach the active centre after NAD has already bound. The structure of the active site and a hypothetical catalytic mechanism are discussed. Sequence homology of FDH with other NAD-dependent formate dehydrogenases and some D-specific dehydrogenases is discussed on the basis of the FDH three-dimensional structure.
About this Structure
2NAC is a Single protein structure of sequence from Pseudomonas sp. with as ligand. Active as Formate dehydrogenase, with EC number 1.2.1.2 Full crystallographic information is available from OCA.
Reference
High resolution structures of holo and apo formate dehydrogenase., Lamzin VS, Dauter Z, Popov VO, Harutyunyan EH, Wilson KS, J Mol Biol. 1994 Feb 25;236(3):759-85. PMID:8114093
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