1e0y: Difference between revisions
New page: left|200px<br /><applet load="1e0y" size="450" color="white" frame="true" align="right" spinBox="true" caption="1e0y, resolution 2.75Å" /> '''STRUCTURE OF THE D17... |
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[[Image:1e0y.jpg|left|200px]]<br /><applet load="1e0y" size=" | [[Image:1e0y.jpg|left|200px]]<br /><applet load="1e0y" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1e0y, resolution 2.75Å" /> | caption="1e0y, resolution 2.75Å" /> | ||
'''STRUCTURE OF THE D170S/T457E DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE'''<br /> | '''STRUCTURE OF THE D170S/T457E DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE'''<br /> | ||
==Overview== | ==Overview== | ||
Vanillyl-alcohol oxidase (VAO) is the prototype of a newly recognized | Vanillyl-alcohol oxidase (VAO) is the prototype of a newly recognized family of structurally related oxidoreductases sharing a conserved FAD-binding domain. The active site of VAO is formed by a cavity where the enzyme is able to catalyze many reactions with phenolic substrates. Among these reactions is the stereospecific hydroxylation of 4-ethylphenol-forming (R)-1-(4'-hydroxyphenyl)ethanol. During this conversion, Asp-170 is probably critical for the hydration of the initially formed p-quinone methide intermediate. By site-directed mutagenesis, the putative active site base has been relocated to the opposite face of the active site cavity. In this way, a change in stereospecificity has been achieved. Like native VAO, the single mutants T457E, D170A, and D170S preferentially converted 4-ethylphenol to the (R)-enantiomer of 1-(4'-hydroxyphenyl)ethanol. The double mutants D170A/T457E and D170S/T457E exhibited an inverted stereospecificity with 4-ethylphenol. Particularly, D170S/T457E was strongly (S)-selective, with an enantiomeric excess of 80%. The crystal structure of D170S/T457E, in complex with trifluoromethylphenol, showed a highly conserved mode of ligand binding and revealed that the distinctive catalytic properties of this mutant are not caused by major structural changes. | ||
==About this Structure== | ==About this Structure== | ||
1E0Y is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with FAD and FCR as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Vanillyl-alcohol_oxidase Vanillyl-alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.38 1.1.3.38] Full crystallographic information is available from [http:// | 1E0Y is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with <scene name='pdbligand=FAD:'>FAD</scene> and <scene name='pdbligand=FCR:'>FCR</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Vanillyl-alcohol_oxidase Vanillyl-alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.38 1.1.3.38] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1E0Y OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Vanillyl-alcohol oxidase]] | [[Category: Vanillyl-alcohol oxidase]] | ||
[[Category: Berkel, W | [[Category: Berkel, W J.H Van.]] | ||
[[Category: Heuvel, R | [[Category: Heuvel, R H.H Van Der.]] | ||
[[Category: Mattevi, A.]] | [[Category: Mattevi, A.]] | ||
[[Category: FAD]] | [[Category: FAD]] | ||
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[[Category: specificity]] | [[Category: specificity]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:22:35 2008'' | ||
Revision as of 10:22, 21 February 2008
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STRUCTURE OF THE D170S/T457E DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE
Overview
Vanillyl-alcohol oxidase (VAO) is the prototype of a newly recognized family of structurally related oxidoreductases sharing a conserved FAD-binding domain. The active site of VAO is formed by a cavity where the enzyme is able to catalyze many reactions with phenolic substrates. Among these reactions is the stereospecific hydroxylation of 4-ethylphenol-forming (R)-1-(4'-hydroxyphenyl)ethanol. During this conversion, Asp-170 is probably critical for the hydration of the initially formed p-quinone methide intermediate. By site-directed mutagenesis, the putative active site base has been relocated to the opposite face of the active site cavity. In this way, a change in stereospecificity has been achieved. Like native VAO, the single mutants T457E, D170A, and D170S preferentially converted 4-ethylphenol to the (R)-enantiomer of 1-(4'-hydroxyphenyl)ethanol. The double mutants D170A/T457E and D170S/T457E exhibited an inverted stereospecificity with 4-ethylphenol. Particularly, D170S/T457E was strongly (S)-selective, with an enantiomeric excess of 80%. The crystal structure of D170S/T457E, in complex with trifluoromethylphenol, showed a highly conserved mode of ligand binding and revealed that the distinctive catalytic properties of this mutant are not caused by major structural changes.
About this Structure
1E0Y is a Single protein structure of sequence from Penicillium simplicissimum with FAD and FCR as ligands. Active as Vanillyl-alcohol oxidase, with EC number 1.1.3.38 Full crystallographic information is available from OCA.
Reference
Inversion of stereospecificity of vanillyl-alcohol oxidase., van Den Heuvel RH, Fraaije MW, Ferrer M, Mattevi A, van Berkel WJ, Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9455-60. PMID:10920192
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