1t2x: Difference between revisions
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[[Image:1t2x.gif|left|200px]] | [[Image:1t2x.gif|left|200px]] | ||
'''Glactose oxidase C383S mutant identified by directed evolution''' | {{Structure | ||
|PDB= 1t2x |SIZE=350|CAPTION= <scene name='initialview01'>1t2x</scene>, resolution 2.3Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=CU:COPPER+(II)+ION'>CU</scene> and <scene name='pdbligand=ACT:ACETATE ION'>ACT</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/Galactose_oxidase Galactose oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.9 1.1.3.9] | |||
|GENE= sac1 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=29916 Fusarium sp.]) | |||
}} | |||
'''Glactose oxidase C383S mutant identified by directed evolution''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
1T2X is a [ | 1T2X is a [[Protein complex]] structure of sequences from [http://en.wikipedia.org/wiki/Fusarium_sp. Fusarium sp.]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1T2X OCA]. | ||
==Reference== | ==Reference== | ||
Structural and kinetic studies of a series of mutants of galactose oxidase identified by directed evolution., Wilkinson D, Akumanyi N, Hurtado-Guerrero R, Dawkes H, Knowles PF, Phillips SE, McPherson MJ, Protein Eng Des Sel. 2004 Feb;17(2):141-8. Epub 2004 Jan 12. PMID:[http:// | Structural and kinetic studies of a series of mutants of galactose oxidase identified by directed evolution., Wilkinson D, Akumanyi N, Hurtado-Guerrero R, Dawkes H, Knowles PF, Phillips SE, McPherson MJ, Protein Eng Des Sel. 2004 Feb;17(2):141-8. Epub 2004 Jan 12. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15047910 15047910] | ||
[[Category: Fusarium sp.]] | [[Category: Fusarium sp.]] | ||
[[Category: Galactose oxidase]] | [[Category: Galactose oxidase]] | ||
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[[Category: mutant form of copper containing enzyme]] | [[Category: mutant form of copper containing enzyme]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 14:13:04 2008'' | ||
Revision as of 15:13, 20 March 2008
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| , resolution 2.3Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | , and | ||||||
| Gene: | sac1 (Fusarium sp.) | ||||||
| Activity: | Galactose oxidase, with EC number 1.1.3.9 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Glactose oxidase C383S mutant identified by directed evolution
OverviewOverview
Galactose oxidase (GO; E.C. 1.1.3.9) is a copper- containing enzyme that oxidizes a range of primary alcohols to aldehydes. This broad substrate specificity is reflected in a high K(M) for substrates. Directed evolution has previously been used to select variants of GO that exhibit enhanced expression and kinetic properties. In assays using unpurified enzyme samples, the variant C383S displayed a 5-fold lower K(M) than wild-type GO. In the present study, we have constructed, expressed, purified and characterized a number of single, double and triple mutants at residues Cys383, Tyr436 and Val494, identified in one of the directed evolution studies, to examine their relative contributions to improved catalytic activity of GO. We report kinetic studies on the various mutant enzymes. In addition, we have determined the three-dimensional structure of the C383S variant. As with many mutations identified in directed evolution experiments, the availability of structural information does not provide a definitive answer to the reason for the improved K(M) in the C383S variant protein.
About this StructureAbout this Structure
1T2X is a Protein complex structure of sequences from Fusarium sp.. Full crystallographic information is available from OCA.
ReferenceReference
Structural and kinetic studies of a series of mutants of galactose oxidase identified by directed evolution., Wilkinson D, Akumanyi N, Hurtado-Guerrero R, Dawkes H, Knowles PF, Phillips SE, McPherson MJ, Protein Eng Des Sel. 2004 Feb;17(2):141-8. Epub 2004 Jan 12. PMID:15047910
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