2M80
Solution structure of yeast dithiol glutaredoxin Grx8
Summary for 2M80
| Entry DOI | 10.2210/pdb2m80/pdb |
| NMR Information | BMRB: 19218 |
| Descriptor | Glutaredoxin-8 (1 entity in total) |
| Functional Keywords | biomolecular, glutaredoxins, glutathione, glutathione disulfide, saccharomyces cerevisiae, oxidation-reduction, tertiary, gsh-dependenet oxidoreductase, electron transport, oxidoreductase |
| Biological source | Saccharomyces cerevisiae (yeast) |
| Cellular location | Cytoplasm: Q05926 |
| Total number of polymer chains | 1 |
| Total formula weight | 13551.50 |
| Authors | |
| Primary citation | Tang, Y.,Zhang, J.,Yu, J.,Xu, L.,Wu, J.,Zhou, C.Z.,Shi, Y. Structure-guided activity enhancement and catalytic mechanism of yeast grx8 Biochemistry, 53:2185-2196, 2014 Cited by PubMed Abstract: 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. PubMed: 24611845DOI: 10.1021/bi401293s PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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