1t2x
From Proteopedia
| Line 1: | Line 1: | ||
[[Image:1t2x.gif|left|200px]] | [[Image:1t2x.gif|left|200px]] | ||
| - | + | <!-- | |
| - | + | The line below this paragraph, containing "STRUCTURE_1t2x", creates the "Structure Box" on the page. | |
| - | + | You may change the PDB parameter (which sets the PDB file loaded into the applet) | |
| - | + | or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | |
| - | + | or leave the SCENE parameter empty for the default display. | |
| - | | | + | --> |
| - | | | + | {{STRUCTURE_1t2x| PDB=1t2x | SCENE= }} |
| - | + | ||
| - | + | ||
| - | }} | + | |
'''Glactose oxidase C383S mutant identified by directed evolution''' | '''Glactose oxidase C383S mutant identified by directed evolution''' | ||
| Line 19: | Line 16: | ||
==About this Structure== | ==About this Structure== | ||
| - | + | 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://www.ncbi.nlm.nih.gov/pubmed/15047910 15047910] | 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: Galactose oxidase]] | [[Category: Galactose oxidase]] | ||
| - | [[Category: Protein complex]] | ||
[[Category: Akumanyi, N.]] | [[Category: Akumanyi, N.]] | ||
[[Category: Dawkes, H.]] | [[Category: Dawkes, H.]] | ||
| Line 34: | Line 29: | ||
[[Category: Wilkinson, D.]] | [[Category: Wilkinson, D.]] | ||
[[Category: 7 blade beta propeller]] | [[Category: 7 blade beta propeller]] | ||
| - | [[Category: | + | [[Category: C383s mutant form]] |
| - | [[Category: | + | [[Category: Mutant form of copper containing enzyme]] |
| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 09:26:58 2008'' | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | |
Revision as of 06:26, 3 May 2008
Glactose oxidase C383S mutant identified by directed evolution
Overview
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 Structure
Full crystallographic information is available from OCA.
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:15047910 Page seeded by OCA on Sat May 3 09:26:58 2008
