From Proteopedia
(Difference between revisions)
proteopedia linkproteopedia link
|
|
| Line 1: |
Line 1: |
| - | [[Image:1usb.gif|left|200px]] | + | {{Seed}} |
| | + | [[Image:1usb.png|left|200px]] |
| | | | |
| | <!-- | | <!-- |
| Line 9: |
Line 10: |
| | {{STRUCTURE_1usb| PDB=1usb | SCENE= }} | | {{STRUCTURE_1usb| PDB=1usb | SCENE= }} |
| | | | |
| - | '''RATIONAL DESIGN OF A NOVEL ENZYME- EFFICIENT THIOESTER HYDROLYSIS ENABLED BY THE INCORPORATION OF A SINGLE HIS RESIDUE INTO HUMAN GLUTATHIONE TRANSFERASE A1-1'''
| + | ===RATIONAL DESIGN OF A NOVEL ENZYME- EFFICIENT THIOESTER HYDROLYSIS ENABLED BY THE INCORPORATION OF A SINGLE HIS RESIDUE INTO HUMAN GLUTATHIONE TRANSFERASE A1-1=== |
| | | | |
| | | | |
| - | ==Overview==
| + | <!-- |
| - | A strategy for rational enzyme design is reported and illustrated by the engineering of a protein catalyst for thiol-ester hydrolysis. Five mutants of human glutathione (GSH; gamma-Glu-Cys-Gly) transferase A1-1 were designed in the search for a catalyst and to provide a set of proteins from which the reaction mechanism could be elucidated. The single mutant A216H catalyzed the hydrolysis of the S-benzoyl ester of GSH under turnover conditions with a k(cat)/K(M) of 156 M(-1) x min(-1), and a catalytic proficiency of >10(7) M(-1) when compared with the first-order rate constant of the uncatalyzed reaction. The wild-type enzyme did not hydrolyze the substrate, and thus, the introduction of a single histidine residue transformed the wild-type enzyme into a turnover system for thiol-ester hydrolysis. By kinetic analysis of single, double, and triple mutants, as well as from studies of reaction products, it was established that the enzyme A216H catalyzes the hydrolysis of the thiol-ester substrate by a mechanism that includes an acyl intermediate at the side chain of Y9. Kinetic measurements and the crystal structure of the A216H GSH complex provided compelling evidence that H216 acts as a general-base catalyst. The introduction of a single His residue into human GSH transferase A1-1 created an unprecedented enzymatic function, suggesting a strategy that may be of broad applicability in the design of new enzymes. The protein catalyst has the hallmarks of a native enzyme and is expected to catalyze various hydrolytic, as well as transesterification, reactions.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_15333749}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 15333749 is the PubMed ID number. |
| | + | --> |
| | + | {{ABSTRACT_PUBMED_15333749}} |
| | | | |
| | ==About this Structure== | | ==About this Structure== |
| Line 27: |
Line 31: |
| | [[Category: Glutathione]] | | [[Category: Glutathione]] |
| | [[Category: Transferase]] | | [[Category: Transferase]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 11:37:15 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Jul 27 14:27:22 2008'' |
Revision as of 11:27, 27 July 2008
Template:STRUCTURE 1usb
RATIONAL DESIGN OF A NOVEL ENZYME- EFFICIENT THIOESTER HYDROLYSIS ENABLED BY THE INCORPORATION OF A SINGLE HIS RESIDUE INTO HUMAN GLUTATHIONE TRANSFERASE A1-1
Template:ABSTRACT PUBMED 15333749
About this Structure
1USB is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
Reference
Incorporation of a single His residue by rational design enables thiol-ester hydrolysis by human glutathione transferase A1-1., Hederos S, Broo KS, Jakobsson E, Kleywegt GJ, Mannervik B, Baltzer L, Proc Natl Acad Sci U S A. 2004 Sep 7;101(36):13163-7. Epub 2004 Aug 27. PMID:15333749
Page seeded by OCA on Sun Jul 27 14:27:22 2008
