3FZA

Crystal structure of poplar glutaredoxin S12 in complex with glutathione and beta-mercaptoethanol

Summary for 3FZA

• Entry DOI: 10.2210/pdb3fza/pdb
• Related: 3FZ9
• Descriptor: Glutaredoxin, BETA-MERCAPTOETHANOL, GLUTATHIONE, ... (4 entities in total)
• Functional Keywords: glutaredoxin, oxidoreductase
• Biological source: Populus tremula x Populus tremuloides
• Total number of polymer chains: 1
• Total formula weight: 12631.44

Authors

Didierjean, C.,Corbier, C.,Koh, C.S.,Rouhier, N.,Jacquot, J.P. (deposition date: 2009-01-24, release date: 2009-02-24, Last modification date: 2024-04-03)

Primary citation

Couturier, J.,Koh, C.S.,Zaffagnini, M.,Winger, A.M.,Gualberto, J.M.,Corbier, C.,Decottignies, P.,Jacquot, J.P.,Lemaire, S.D.,Didierjean, C.,Rouhier, N.
Structure-function relationship of the chloroplastic glutaredoxin S12 with an atypical WCSYS active site.
J.Biol.Chem., 284:9299-9310, 2009

PubMed Abstract: Glutaredoxins (Grxs) are efficient catalysts for the reduction of mixed disulfides in glutathionylated proteins, using glutathione or thioredoxin reductases for their regeneration. Using GFP fusion, we have shown that poplar GrxS12, which possesses a monothiol (28)WCSYS(32) active site, is localized in chloroplasts. In the presence of reduced glutathione, the recombinant protein is able to reduce in vitro substrates, such as hydroxyethyldisulfide and dehydroascorbate, and to regenerate the glutathionylated glyceraldehyde-3-phosphate dehydrogenase. Although the protein possesses two conserved cysteines, it is functioning through a monothiol mechanism, the conserved C terminus cysteine (Cys(87)) being dispensable, since the C87S variant is fully active in all activity assays. Biochemical and crystallographic studies revealed that Cys(87) exhibits a certain reactivity, since its pK(a) is around 5.6. Coupled with thiol titration, fluorescence, and mass spectrometry analyses, the resolution of poplar GrxS12 x-ray crystal structure shows that the only oxidation state is a glutathionylated derivative of the active site cysteine (Cys(29)) and that the enzyme does not form inter- or intramolecular disulfides. Contrary to some plant Grxs, GrxS12 does not incorporate an iron-sulfur cluster in its wild-type form, but when the active site is mutated into YCSYS, it binds a [2Fe-2S] cluster, indicating that the single Trp residue prevents this incorporation.

PubMed: 19158074
DOI: 10.1074/jbc.M807998200

Experimental method

X-RAY DIFFRACTION (1.8 Å)