1usb

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[[Image:1usb.gif|left|200px]]<br /><applet load="1usb" size="350" color="white" frame="true" align="right" spinBox="true"
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[[Image:1usb.gif|left|200px]]
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caption="1usb, resolution 2.07&Aring;" />
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'''RATIONAL DESIGN OF A NOVEL ENZYME- EFFICIENT THIOESTER HYDROLYSIS ENABLED BY THE INCORPORATION OF A SINGLE HIS RESIDUE INTO HUMAN GLUTATHIONE TRANSFERASE A1-1'''<br />
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{{Structure
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|PDB= 1usb |SIZE=350|CAPTION= <scene name='initialview01'>1usb</scene>, resolution 2.07&Aring;
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|SITE= <scene name='pdbsite=AC1:K+Binding+Site+For+Chain+B'>AC1</scene>
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|LIGAND= <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene> and <scene name='pdbligand=GSH:GLUTATHIONE'>GSH</scene>
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|ACTIVITY= [http://en.wikipedia.org/wiki/Glutathione_transferase Glutathione transferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.5.1.18 2.5.1.18]
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|GENE=
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}}
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'''RATIONAL DESIGN OF A NOVEL ENZYME- EFFICIENT THIOESTER HYDROLYSIS ENABLED BY THE INCORPORATION OF A SINGLE HIS RESIDUE INTO HUMAN GLUTATHIONE TRANSFERASE A1-1'''
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==Overview==
==Overview==
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==About this Structure==
==About this Structure==
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1USB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=CL:'>CL</scene>, <scene name='pdbligand=K:'>K</scene> and <scene name='pdbligand=GSH:'>GSH</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glutathione_transferase Glutathione transferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.5.1.18 2.5.1.18] Known structural/functional Site: <scene name='pdbsite=AC1:K+Binding+Site+For+Chain+B'>AC1</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1USB OCA].
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1USB is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1USB OCA].
==Reference==
==Reference==
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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:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=15333749 15333749]
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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:[http://www.ncbi.nlm.nih.gov/pubmed/15333749 15333749]
[[Category: Glutathione transferase]]
[[Category: Glutathione transferase]]
[[Category: Homo sapiens]]
[[Category: Homo sapiens]]
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[[Category: transferase]]
[[Category: transferase]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:27:48 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 14:36:01 2008''

Revision as of 12:36, 20 March 2008

Image:1usb.gif


PDB ID 1usb

Drag the structure with the mouse to rotate
, resolution 2.07Å
Sites:
Ligands: , and
Activity: Glutathione transferase, with EC number 2.5.1.18
Coordinates: save as pdb, mmCIF, xml



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.

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

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