TY  - JOUR
AU  - Liedgens, Linda
AU  - Zimmermann, Jannik
AU  - Wäschenbach, Lucas
AU  - Geissel, Fabian
AU  - Laporte, Hugo
AU  - Gohlke, Holger
AU  - Morgan, Bruce
AU  - Deponte, Marcel
TI  - Quantitative assessment of the determinant structural differences between redox-active and inactive glutaredoxins
JO  - Nature Communications
VL  - 11
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Nature Publishing Group UK
M1  - FZJ-2020-01704
SP  - 1725
PY  - 2020
AB  - Class I glutaredoxins are enzymatically active, glutathione-dependent oxidoreductases, whilst class II glutaredoxins are typically enzymatically inactive, Fe-S cluster-binding proteins. Enzymatically active glutaredoxins harbor both a glutathione-scaffold site for reacting with glutathionylated disulfide substrates and a glutathione-activator site for reacting with reduced glutathione. Here, using yeast ScGrx7 as a model protein, we comprehensively identified and characterized key residues from four distinct protein regions, as well as the covalently bound glutathione moiety, and quantified their contribution to both interaction sites. Additionally, we developed a redox-sensitive GFP2-based assay, which allowed the real-time assessment of glutaredoxin structure-function relationships inside living cells. Finally, we employed this assay to rapidly screen multiple glutaredoxin mutants, ultimately enabling us to convert enzymatically active and inactive glutaredoxins into each other. In summary, we have gained a comprehensive understanding of the mechanistic underpinnings of glutaredoxin catalysis and have elucidated the determinant structural differences between the two main classes of glutaredoxins.
LB  - PUB:(DE-HGF)16
C6  - pmid:32265442
UR  - <Go to ISI:>//WOS:000526532500003
DO  - DOI:10.1038/s41467-020-15441-3
UR  - https://juser.fz-juelich.de/record/874920
ER  -