A novel glutaredoxin domain-containing peroxiredoxin 'AII1541' protects the N-2-fixing cyanobacterium Anabaena PCC 7120 from oxidative stress

作者：Banerjee Manisha; Ballal Anand; Apte Shree K^*

来源：Biochemical Journal, 2012, 442: 671-680.

DOI：10.1042/BJ20111877

• AII1541
• Anabaena
• antioxidant
• peroxiredoxin-glutaredoxin hybrid
• reaction mechanism
• stress tolerance

Prxs (peroxiredoxins) are ubiquitous thiol-based peroxidases that detoxify toxic peroxides. The Anabaena PCC 7120 genome harbours seven genes/ORFs (open reading frames) which have homology with Prxs. One of these (aII1541) was identified to encode a novel Grx (glutaredoxin) domain-containing Prx by bioinformatic analysis. A recombinant N-terminal histidine-tagged AII1541 protein was overexpressed in Escherichia coli and purified. Analysis with the protein alkylating agent AMS (4-acetamido-4'-maleimidyl-stilbene-2,2'-disulfonate) showed AII1541 to form an intra-molecular disulfide bond. The AII1541 protein used glutathione (GSH) more efficiently than Trx (thioredoxin) to detoxify H2O2. Deletion of the Grx domain from AII1541 resulted in loss of GSH-dependent peroxidase activity. Employing site-directed mutagenesis, the cysteine residues at positions 50 and 75 were identified as peroxidatic and resolving cysteine residues respectively, whereas both the cysteine residues within the Grx domain (positions 181 and 184) were shown to be essential for GSH-dependent peroxidase activity. On the basis of these data, a reaction mechanism has been proposed for AII1541. In vitro AII1541 protein protected plasmid DNA from oxidative damage. In Anabaena PCC 7120, aII1541 was transcriptionally activated under oxidative stress. Recombinant Anabaena PCC 7120 strain overexpressing AII1541 protein showed superior oxidative stress tolerance to H2O2 as compared with the wild-type strain. The results suggest that the glutathione-dependent peroxidase AII1541 plays an important role in protecting Anabaena from oxidative stress.

• 出版日期2012-3-15