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| | ==Chimeric GST== | | ==Chimeric GST== |
| - | <StructureSection load='5ld0' size='340' side='right' caption='[[5ld0]], [[Resolution|resolution]] 1.60Å' scene=''> | + | <StructureSection load='5ld0' size='340' side='right'caption='[[5ld0]], [[Resolution|resolution]] 1.60Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5ld0]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5LD0 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5LD0 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5ld0]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] and [https://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5LD0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5LD0 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.6Å</td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5ld0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ld0 OCA], [http://pdbe.org/5ld0 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5ld0 RCSB], [http://www.ebi.ac.uk/pdbsum/5ld0 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5ld0 ProSAT]</span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene></td></tr> |
| | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5ld0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ld0 OCA], [https://pdbe.org/5ld0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5ld0 RCSB], [https://www.ebi.ac.uk/pdbsum/5ld0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5ld0 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/GSTA1_HUMAN GSTA1_HUMAN]] Conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles.<ref>PMID:20606271</ref> | + | [https://www.uniprot.org/uniprot/GSTA2_RAT GSTA2_RAT] Catalyzes the conjugation of glutathione to a large variety of electrophilic compounds.[UniProtKB:P10648][https://www.uniprot.org/uniprot/GSTA1_HUMAN GSTA1_HUMAN] Conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles.<ref>PMID:20606271</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 5ld0" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 5ld0" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Glutathione S-transferase 3D structures|Glutathione S-transferase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Axarli, A]] | + | [[Category: Homo sapiens]] |
| - | [[Category: Chronopoulou, E G]] | + | [[Category: Large Structures]] |
| - | [[Category: Labrou, N E]] | + | [[Category: Rattus norvegicus]] |
| - | [[Category: Muleta, A W]] | + | [[Category: Axarli A]] |
| - | [[Category: Papageorgiou, A C]] | + | [[Category: Chronopoulou EG]] |
| - | [[Category: Directed evolution]] | + | [[Category: Labrou NE]] |
| - | [[Category: Glutathione transferase a1-1]] | + | [[Category: Muleta AW]] |
| - | [[Category: Protein stability]] | + | [[Category: Papageorgiou AC]] |
| - | [[Category: Transferase]]
| + | |
| Structural highlights
Function
GSTA2_RAT Catalyzes the conjugation of glutathione to a large variety of electrophilic compounds.[UniProtKB:P10648]GSTA1_HUMAN Conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles.[1]
Publication Abstract from PubMed
BACKGROUND: Glutathione transferases (GSTs) are a family of detoxification enzymes that catalyse the conjugation of glutathione (GSH) to electrophilic compounds. METHODS: A library of alpha class GSTs was constructed by DNA shuffling using the DNA encoding the human glutathione transferase A1-1 (hGSTA1-1) and the rat glutathione transferase A1-1 (rGSTA1-1). RESULTS: Activity screening of the library allowed the selection of a chimeric enzyme variant (GSTD4) that displayed high affinity towards GSH and GSH-Sepharose affinity adsorbent, higher kcat/Km and improved thermal stability, compared to the parent enzymes. The crystal structures of the GSTD4 enzyme in free form and in complex with GSH were determined to 1.6A and 2.3A resolution, respectively. Analysis of the GSTD4 structure showed subtle conformational changes in the GSH-binding site and in electron-sharing network that may contribute to the increased GSH affinity. The shuffled variant GSTD4 was further optimised for improved oxidative stability employing site-saturation mutagenesis. The Cys112Ser mutation confers optimal oxidative stability and kinetic properties in the GSTD4 enzyme. CONCLUSIONS: DNA shuffling allowed the creation of a chimeric enzyme variant with improved properties, compared to the parent enzymes. X-ray crystallography shed light on how recombination of a specific segments from homologous GSTA1-1 together with point mutations gives rise to a new functionally competent enzyme with improved binding, catalytic properties and stability. GENERAL SIGNIFICANCE: Such an engineered GST would be useful in biotechnology as affinity tool in affinity chromatography as well as a biocatalytic matrix for the construction of biochips or enzyme biosensors.
Directed Evolution of Glutathione Transferases Towards a Selective Glutathione-Binding Site and Improved Oxidative Stability.,Axarli I, Muleta AW, Chronopoulou EG, Papageorgiou AC, Labrou NE Biochim Biophys Acta. 2016 Sep 6. pii: S0304-4165(16)30326-9. doi:, 10.1016/j.bbagen.2016.09.004. PMID:27612661[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Achilonu I, Gildenhuys S, Fisher L, Burke J, Fanucchi S, Sewell BT, Fernandes M, Dirr HW. The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jul 1;66(Pt, 7):776-80. Epub 2010 Jun 23. PMID:20606271 doi:10.1107/S1744309110019135
- ↑ Axarli I, Muleta AW, Chronopoulou EG, Papageorgiou AC, Labrou NE. Directed Evolution of Glutathione Transferases Towards a Selective Glutathione-Binding Site and Improved Oxidative Stability. Biochim Biophys Acta. 2016 Sep 6. pii: S0304-4165(16)30326-9. doi:, 10.1016/j.bbagen.2016.09.004. PMID:27612661 doi:http://dx.doi.org/10.1016/j.bbagen.2016.09.004
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