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| | ==Solution structure of yeast dithiol glutaredoxin Grx8== | | ==Solution structure of yeast dithiol glutaredoxin Grx8== |
| - | <StructureSection load='2m80' size='340' side='right'caption='[[2m80]], [[NMR_Ensembles_of_Models | 20 NMR models]]' scene=''> | + | <StructureSection load='2m80' size='340' side='right'caption='[[2m80]]' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[2m80]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Baker's_yeast Baker's yeast]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2M80 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2M80 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[2m80]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Saccharomyces_cerevisiae_S288C Saccharomyces cerevisiae S288C]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2M80 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2M80 FirstGlance]. <br> |
| - | </td></tr><tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">GRX8 ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=559292 Baker's yeast])</td></tr> | + | </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=2m80 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2m80 OCA], [https://pdbe.org/2m80 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2m80 RCSB], [https://www.ebi.ac.uk/pdbsum/2m80 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2m80 ProSAT]</span></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=2m80 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2m80 OCA], [https://pdbe.org/2m80 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2m80 RCSB], [https://www.ebi.ac.uk/pdbsum/2m80 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2m80 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[https://www.uniprot.org/uniprot/GLRX8_YEAST GLRX8_YEAST]] Glutathione-dependent oxidoreductase with lower activity compared to the other members of the glutaredoxin family. The disulfide bond functions as an electron carrier in the glutathione-dependent synthesis of deoxyribonucleotides by the enzyme ribonucleotide reductase.<ref>PMID:18400945</ref> <ref>PMID:19166312</ref>
| + | [https://www.uniprot.org/uniprot/GLRX8_YEAST GLRX8_YEAST] Glutathione-dependent oxidoreductase with lower activity compared to the other members of the glutaredoxin family. The disulfide bond functions as an electron carrier in the glutathione-dependent synthesis of deoxyribonucleotides by the enzyme ribonucleotide reductase.<ref>PMID:18400945</ref> <ref>PMID:19166312</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Baker's yeast]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Shi, Y]] | + | [[Category: Saccharomyces cerevisiae S288C]] |
| - | [[Category: Tang, Y]] | + | [[Category: Shi Y]] |
| - | [[Category: Wu, J]] | + | [[Category: Tang Y]] |
| - | [[Category: Yu, J]] | + | [[Category: Wu J]] |
| - | [[Category: Zhang, J]] | + | [[Category: Yu J]] |
| - | [[Category: Zhou, C Z]] | + | [[Category: Zhang J]] |
| - | [[Category: Biomolecular]]
| + | [[Category: Zhou CZ]] |
| - | [[Category: Electron transport]]
| + | |
| - | [[Category: Glutaredoxin]]
| + | |
| - | [[Category: Glutathione]]
| + | |
| - | [[Category: Glutathione disulfide]]
| + | |
| - | [[Category: Gsh-dependenet oxidoreductase]]
| + | |
| - | [[Category: Oxidation-reduction]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| - | [[Category: Saccharomyces cerevisiae]]
| + | |
| - | [[Category: Tertiary]]
| + | |
| Structural highlights
Function
GLRX8_YEAST Glutathione-dependent oxidoreductase with lower activity compared to the other members of the glutaredoxin family. The disulfide bond functions as an electron carrier in the glutathione-dependent synthesis of deoxyribonucleotides by the enzyme ribonucleotide reductase.[1] [2]
Publication Abstract from PubMed
Glutaredoxins (Grxs) are wide-spread oxidoreductases that are found in all kingdoms of life. The yeast Saccharomyces cerevisiae encodes eight Grxs, among which, Grx8 shares a sequence identity of 30 and 23% with typical dithiol Grx1 and Grx2, respectively, but it exhibits a much lower GSH-dependent oxidoreductase activity. To elucidate its catalytic mechanism, we solved the solution structure of Grx8, which displays a typical Grx fold. Structural analysis indicated that Grx8 possesses a negatively charged CXXC motif (Cys(33)-Pro(34)-Asp(35)-Cys(36)) and a GSH-recognition site, which are distinct from Grx1 and Grx2. Subsequent structure-guided site mutations revealed that the D35Y single mutant and N80T/L81V double mutant possess increased activity of 10- and 11-fold, respectively; moreover, the D35Y/N80T/L81V triple mutant has increased activity of up to 44-fold, which is comparable to that of canonical Grx. Biochemical analyses suggested that the increase in catalytic efficiency resulted from a decreased pKa value of catalytic cysteine Cys33 and/or enhancement of the putative GSH-recognition site. Moreover, NMR chemical shift perturbation analyses combined with GSH analogue inhibition assays enabled us to elucidate that wild-type Grx8 and all mutants adopt a ping-pong mechanism of catalysis. All together, these findings provide structural insights into the catalytic mechanism of dithiol Grxs.
Structure-guided activity enhancement and catalytic mechanism of yeast grx8.,Tang Y, Zhang J, Yu J, Xu L, Wu J, Zhou CZ, Shi Y Biochemistry. 2014 Apr 8;53(13):2185-96. doi: 10.1021/bi401293s. Epub 2014 Mar, 25. PMID:24611845[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Mesecke N, Spang A, Deponte M, Herrmann JM. A novel group of glutaredoxins in the cis-Golgi critical for oxidative stress resistance. Mol Biol Cell. 2008 Jun;19(6):2673-80. doi: 10.1091/mbc.E07-09-0896. Epub 2008, Apr 9. PMID:18400945 doi:http://dx.doi.org/10.1091/mbc.E07-09-0896
- ↑ Eckers E, Bien M, Stroobant V, Herrmann JM, Deponte M. Biochemical characterization of dithiol glutaredoxin 8 from Saccharomyces cerevisiae: the catalytic redox mechanism redux. Biochemistry. 2009 Feb 17;48(6):1410-23. doi: 10.1021/bi801859b. PMID:19166312 doi:http://dx.doi.org/10.1021/bi801859b
- ↑ Tang Y, Zhang J, Yu J, Xu L, Wu J, Zhou CZ, Shi Y. Structure-guided activity enhancement and catalytic mechanism of yeast grx8. Biochemistry. 2014 Apr 8;53(13):2185-96. doi: 10.1021/bi401293s. Epub 2014 Mar, 25. PMID:24611845 doi:http://dx.doi.org/10.1021/bi401293s
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