Glutaredoxins employ parallel monothiol-dithiol mechanisms to catalyze thiol-disulfide exchanges with protein disulfides
Ashwinie A Ukuwela, Ashley I Bush, Anthony G Wedd, Zhiguang Xiao
CHEMICAL SCIENCE | ROYAL SOC CHEMISTRY | Published : 2018
Glutaredoxins (Grxs) are a family of glutathione (GSH)-dependent thiol-disulfide oxidoreductases. They feature GSH-binding sites that directly connect the reversible redox chemistry of protein thiols to the abundant cellular nonprotein thiol pool GSSG/GSH. This work studied the pathways for oxidation of protein dithiols P(SH)2 and reduction of protein disulfides P(SS) catalyzed by Homo sapiens HsGrx1 and Escherichia coli EcGrx1. The metal-binding domain HMA4n(SH)2 was chosen as substrate as it contains a solvent-exposed CysCys motif. Quenching of the reactions with excess iodoacetamide followed by protein speciation analysis via ESI-MS allowed interception and characterization of both substr..View full abstract
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Awarded by Australian Research Council
This work was supported by funds from the Australian Research Council Grant DP130100728. Additional support for AAU was generously provided by the Norma Hilda Schuster (nee Swift) Scholarship Fund. ESI-MS analysis was conducted at the Bio21 Mass Spectrometry and Proteomics Facility.