Structural basis for the different activities of yeast Grx1 and Grx2.
Yeast glutaredoxins Grx1 and Grx2 catalyze the reduction of both inter- and intra-molecular disulfide bonds using glutathione (GSH) as the electron donor. Although sharing the same dithiolic CPYC active site and a sequence identity of 64%, they have been proved to play different roles during oxidative stress and to possess different glutathione-disulfide reductase activities. To address the structural basis of these differences, we solved the crystal structures of Grx2 in oxidized and reduced forms, at 2.10 A and 1.50 A, respectively. With the Grx1 structures we previously reported, comparative structural analyses revealed that Grx1 and Grx2 share a similar GSH binding site, except for a single residue substitution from Asp89 in Grx1 to Ser123 in Grx2. Site-directed mutagenesis in combination with activity assays further proved this single residue variation is critical for the different activities of yeast Grx1 and Grx2.
Li
Wei-Fang
WF
Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei Anhui 230027, PR China.
Yu
Jiang
J
Ma
Xiao-Xiao
XX
Teng
Yan-Bin
YB
Luo
Ming
M
Tang
Ya-Jun
YJ
Zhou
Cong-Zhao
CZ
D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D054477:Glutaredoxins / Q000737:chemistry* / Q000502:physiology*; D005978:Glutathione / Q000737:chemistry; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011485:Protein Binding; D011487:Protein Conformation; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000502:physiology*; D017386:Sequence Homology, Amino Acid
D054477:Glutaredoxins; C530831:Grx1 protein, S cerevisiae; C527885:Grx2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D005978:Glutathione