From Proteopedia
(Difference between revisions)
proteopedia linkproteopedia link
|
|
| Line 1: |
Line 1: |
| - | [[Image:1toe.gif|left|200px]] | + | {{Seed}} |
| | + | [[Image:1toe.png|left|200px]] |
| | | | |
| | <!-- | | <!-- |
| Line 9: |
Line 10: |
| | {{STRUCTURE_1toe| PDB=1toe | SCENE= }} | | {{STRUCTURE_1toe| PDB=1toe | SCENE= }} |
| | | | |
| - | '''Unliganded structure of Hexamutant + A293D mutant of E. coli aspartate aminotransferase'''
| + | ===Unliganded structure of Hexamutant + A293D mutant of E. coli aspartate aminotransferase=== |
| | | | |
| | | | |
| - | ==Overview==
| + | <!-- |
| - | Several mutant Escherichia coli aspartate aminotransferases (eAATases) have been characterized in the attempt to evolve or rationally redesign the substrate specificity of eAATase into that of E. coli tyrosine aminotransferase (eTATase). These include HEX (designed), HEX + A293D (design followed by directed evolution), and SRHEPT (directed evolution). The A293D mutation realized from directed evolution of HEX is here imported into the SRHEPT platform by site-directed mutagenesis, resulting in an enzyme (SRHEPT + A293D) with nearly the same ratio of k(cat)/K(m)(Phe) to k(cat)/K(m)(Asp) as that of wild-type eTATase. The A293D substitution is an important specificity determinant; it selectively disfavors interactions with dicarboxylic substrates and inhibitors compared to aromatic ones. Context dependence analysis is generalized to provide quantitative comparisons of a common substitution in two or more different protein scaffolds. High-resolution crystal structures of ligand complexes of HEX + A293D, SRHEPT, and SRHEPT + A293D were determined. We find that in both SRHEPT + A293D and HEX + A293D, the additional mutation holds the Arg 292 side chain away from the active site to allow increased specificity for phenylalanine over aspartate. The resulting movement of Arg 292 allows greater flexibility of the small domain in HEX + A293D. While HEX is always in the closed conformation, HEX + A293D is observed in both the closed and a novel open conformation, allowing for more rapid product release.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_15461450}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 15461450 is the PubMed ID number. |
| | + | --> |
| | + | {{ABSTRACT_PUBMED_15461450}} |
| | | | |
| | ==About this Structure== | | ==About this Structure== |
| Line 29: |
Line 33: |
| | [[Category: McElroy, K E.]] | | [[Category: McElroy, K E.]] |
| | [[Category: Aspartate aminotransferase hexamutant]] | | [[Category: Aspartate aminotransferase hexamutant]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 10:11:06 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 18:08:17 2008'' |
Revision as of 15:08, 28 July 2008
Template:STRUCTURE 1toe
Unliganded structure of Hexamutant + A293D mutant of E. coli aspartate aminotransferase
Template:ABSTRACT PUBMED 15461450
About this Structure
1TOE is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
Reference
Narrowing substrate specificity in a directly evolved enzyme: the A293D mutant of aspartate aminotransferase., Chow MA, McElroy KE, Corbett KD, Berger JM, Kirsch JF, Biochemistry. 2004 Oct 12;43(40):12780-7. PMID:15461450
Page seeded by OCA on Mon Jul 28 18:08:17 2008
