Scheda di dettaglio – i prodotti della ricerca
| Dato | Valore |
|---|---|
| Title | Involvement of the Azotobacter vinelandii Rhodanese-like protein RhdA in the glutathione regeneration pathway |
| Abstract | The phenotypic features of the Azotobacter vinelandii RhdA mutant MV474 (in which the rhdA gene was deleted) indicated that defects in antioxidant systems in this organism were related to the expression of the tandem-domain rhodanese RhdA. In this work, further insights on the effects of the oxidative imbalance generated by the absence of RhdA (e.g. increased levels of lipid hydroperoxides) are provided. Starting from the evidence that glutathione was depleted in MV474, and using both in silico and in vitro approaches, here we studied the interaction of wild-type RhdA and Cys(230) Ala site-directed RhdA mutant with glutathione species. We found that RhdA was able to bind in vitro reduced glutathione (GSH) and that RhdA-Cys(230) residue was mandatory for the complex formation. RhdA catalyzed glutathione-disulfide formation in the presence of a system generating the glutathione thiyl radical (GS(.), an oxidized form of GSH), thereby facilitating GSH regeneration. This reaction was negligible when the Cys230 Ala RhdA mutant was used. The efficiency of RhdA as catalyst in GS(.)-scavenging activity is discussed on the basis of the measured parameters of both interaction with glutathione species and kinetic studies. |
| Source | PloS one 7 (9), pp. e0045193 |
| Keywords | RhdARedox homeostasisGlutathioneGlutathione-thiyl radicalLipid hydroperoxidesAzotobacter vinelandii |
| Journal | PloS one |
| Editor | Public Library of Science, San Francisco, CA, Stati Uniti d'America |
| Year | 2012 |
| Type | Articolo in rivista |
| DOI | 10.1371/journal.pone.0045193 |
| Authors | William Remelli; Nicoletta Guerrieri; Jennifer Klodmann; Jutta Papenbrock; Silvia Pagani; Fabio Forlani |
| Text | 323668 2012 10.1371/journal.pone.0045193 ISI Web of Science WOS 000309556100031 RhdA Redox homeostasis Glutathione Glutathione thiyl radical Lipid hydroperoxides Azotobacter vinelandii Involvement of the Azotobacter vinelandii Rhodanese like protein RhdA in the glutathione regeneration pathway William Remelli; Nicoletta Guerrieri; Jennifer Klodmann; Jutta Papenbrock; Silvia Pagani; Fabio Forlani University of Milan; University of Hannover; University of Hannover The phenotypic features of the Azotobacter vinelandii RhdA mutant MV474 in which the rhdA gene was deleted indicated that defects in antioxidant systems in this organism were related to the expression of the tandem domain rhodanese RhdA. In this work, further insights on the effects of the oxidative imbalance generated by the absence of RhdA e.g. increased levels of lipid hydroperoxides are provided. Starting from the evidence that glutathione was depleted in MV474, and using both in silico and in vitro approaches, here we studied the interaction of wild type RhdA and Cys 230 Ala site directed RhdA mutant with glutathione species. We found that RhdA was able to bind in vitro reduced glutathione GSH and that RhdA Cys 230 residue was mandatory for the complex formation. RhdA catalyzed glutathione disulfide formation in the presence of a system generating the glutathione thiyl radical GS . , an oxidized form of GSH , thereby facilitating GSH regeneration. This reaction was negligible when the Cys230 Ala RhdA mutant was used. The efficiency of RhdA as catalyst in GS . scavenging activity is discussed on the basis of the measured parameters of both interaction with glutathione species and kinetic studies. 7 Published version Articolo 2012_PLOS_ONE_7_9_e45193.pdf Articolo in rivista Public Library of Science 1932 6203 PloS one PloS one PLoS ONE PloS one Public Library of Science one PLoS 1 nicoletta.guerrieri GUERRIERI NICOLETTA TA.P02.014.002 Impatto dei cambiamenti globali sugli ecosistemi acquatici |