2vfs
From Proteopedia
| Line 4: | Line 4: | ||
==Overview== | ==Overview== | ||
| - | Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble | + | Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex with both six-carbon (mannitol and sorbitol) and five-carbon substrates (xylitol). AldO shares the same folding topology of the members of the vanillyl-alcohol oxidase family of flavoenzymes and exhibits a covalently linked FAD which is located at the bottom of a funnel-shaped pocket that forms the active site. The high resolution of the three-dimensional structures highlights a well-defined hydrogen-bonding network that tightly constrains the substrate in the productive conformation for catalysis. Substrate binding occurs through a lock-and-key mechanism and does not induce conformational changes with respect to the ligand-free protein. A network of charged residues is proposed to favor catalysis through stabilization of the deprotonated form of the substrate. A His side chain acts as back door that "pushes" the substrate-reactive carbon atom toward the N5-C4a locus of the flavin. Analysis of the three-dimensional structure reveals possible pathways for diffusion of molecular oxygen and a small cavity on the re side of the flavin that may host oxygen during FAD reoxidation. These features combined with the tight shape of the catalytic site provide insights into the mechanism of AldO-mediated regioselective oxidation reactions and its substrate specificity. |
==About this Structure== | ==About this Structure== | ||
| - | 2VFS is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_coelicolor Streptomyces coelicolor] with <scene name='pdbligand=XYL:'>XYL</scene>, <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=FAD:'>FAD</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Xylitol_oxidase Xylitol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.41 1.1.3.41] Known structural/functional Sites: <scene name='pdbsite=AC1:Fad Binding Site For Chain A'>AC1</scene>, <scene name='pdbsite=AC2:Xyl Binding Site For Chain A'>AC2</scene> and <scene name='pdbsite=AC3:Cl Binding Site For Chain A'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VFS OCA]. | + | 2VFS is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_coelicolor Streptomyces coelicolor] with <scene name='pdbligand=XYL:'>XYL</scene>, <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=FAD:'>FAD</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Xylitol_oxidase Xylitol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.41 1.1.3.41] Known structural/functional Sites: <scene name='pdbsite=AC1:Fad+Binding+Site+For+Chain+A'>AC1</scene>, <scene name='pdbsite=AC2:Xyl+Binding+Site+For+Chain+A'>AC2</scene> and <scene name='pdbsite=AC3:Cl+Binding+Site+For+Chain+A'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VFS OCA]. |
==Reference== | ==Reference== | ||
| - | Structural | + | Structural analysis of the catalytic mechanism and stereoselectivity in Streptomyces coelicolor alditol oxidase., Forneris F, Heuts DP, Delvecchio M, Rovida S, Fraaije MW, Mattevi A, Biochemistry. 2008 Jan 22;47(3):978-85. Epub 2007 Dec 23. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18154360 18154360] |
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Streptomyces coelicolor]] | [[Category: Streptomyces coelicolor]] | ||
| Line 27: | Line 27: | ||
[[Category: sugar]] | [[Category: sugar]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:55:29 2008'' |
Revision as of 16:55, 21 February 2008
|
ALDITOL OXIDASE FROM STREPTOMYCES COELICOLOR A3(2): COMPLEX WITH XYLITOL
Overview
Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex with both six-carbon (mannitol and sorbitol) and five-carbon substrates (xylitol). AldO shares the same folding topology of the members of the vanillyl-alcohol oxidase family of flavoenzymes and exhibits a covalently linked FAD which is located at the bottom of a funnel-shaped pocket that forms the active site. The high resolution of the three-dimensional structures highlights a well-defined hydrogen-bonding network that tightly constrains the substrate in the productive conformation for catalysis. Substrate binding occurs through a lock-and-key mechanism and does not induce conformational changes with respect to the ligand-free protein. A network of charged residues is proposed to favor catalysis through stabilization of the deprotonated form of the substrate. A His side chain acts as back door that "pushes" the substrate-reactive carbon atom toward the N5-C4a locus of the flavin. Analysis of the three-dimensional structure reveals possible pathways for diffusion of molecular oxygen and a small cavity on the re side of the flavin that may host oxygen during FAD reoxidation. These features combined with the tight shape of the catalytic site provide insights into the mechanism of AldO-mediated regioselective oxidation reactions and its substrate specificity.
About this Structure
2VFS is a Single protein structure of sequence from Streptomyces coelicolor with , and as ligands. Active as Xylitol oxidase, with EC number 1.1.3.41 Known structural/functional Sites: , and . Full crystallographic information is available from OCA.
Reference
Structural analysis of the catalytic mechanism and stereoselectivity in Streptomyces coelicolor alditol oxidase., Forneris F, Heuts DP, Delvecchio M, Rovida S, Fraaije MW, Mattevi A, Biochemistry. 2008 Jan 22;47(3):978-85. Epub 2007 Dec 23. PMID:18154360
Page seeded by OCA on Thu Feb 21 18:55:29 2008
Categories: Single protein | Streptomyces coelicolor | Xylitol oxidase | Forneris, F. | Mattevi, A. | CL | FAD | XYL | Fad | Flavin | Flavoprotein | Oxidase | Oxidoreductase | Polyol | Sugar
