2m80: Difference between revisions
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''' | ==Solution structure of yeast dithiol glutaredoxin Grx8== | ||
<StructureSection load='2m80' size='340' side='right' caption='[[2m80]], [[NMR_Ensembles_of_Models | 20 NMR models]]' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[2m80]] is a 1 chain structure. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2M80 OCA]. <br> | |||
</td></tr><tr><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Glucokinase Glucokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.2 2.7.1.2] </span></td></tr> | |||
<tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2m80 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2m80 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=2m80 RCSB], [http://www.ebi.ac.uk/pdbsum/2m80 PDBsum]</span></td></tr> | |||
<table> | |||
<div style="background-color:#fffaf0;"> | |||
== 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<ref>PMID:24611845</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Shi, Y.]] | |||
[[Category: Tang, Y.]] | |||
[[Category: Wu, J.]] | |||
[[Category: Yu, J.]] | |||
[[Category: Zhang, J.]] | |||
[[Category: Zhou, C Z.]] | |||
[[Category: Biomolecular]] | |||
[[Category: Electron transport]] | |||
[[Category: Glutaredoxin]] | |||
[[Category: Glutathione]] | |||
[[Category: Glutathione disulfide]] | |||
[[Category: Gsh-dependenet oxidoreductase]] | |||
[[Category: Oxidation-reduction]] | |||
[[Category: Oxidoreductase]] | |||
[[Category: Saccharomyces cerevisiae]] | |||
[[Category: Tertiary]] | |||
Revision as of 08:12, 7 May 2014
Solution structure of yeast dithiol glutaredoxin Grx8
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