Basal Glutathionylation of Na,K-ATPase α-Subunit Depends on Redox Status of Cells during the Enzyme Biosynthesis
Table 1
Structures of Na,K-ATPase from pig kidney with resolution better than 4 .
PDB ID
3B8E
3KDP
3WGU
3WGV
4HYT
Resolution
3.50
3.50
2.80
2.80
3.40
Species, organ
Pig kidney
Pig kidney
Pig kidney
Pig kidney
Pig kidney
Conformation
E2P
—
E1P
E1P with oligomycin
E2P with ouabain
Method
Vapor diffusion and hanging drop
Vapor diffusion
Vapor diffusion and hanging drop
Vapor diffusion and hanging drop
Hanging drop
pH
7.0
7.0
6.2
6.2
6.2
Temperature
292.0 K
292.0 K
283.0 K
283.0 K
292.0 K
Details
14% PEG 2000 mme, 0.2 M choline chloride, 4% glycerol, 4% MPD, 0.04 M DTT, and 0.1–0.4% beta-DDM
14% PEG 2000 mme, 0.2 M choline chloride, 4% glycerol, 4% MPD, 0.04 M DTT, and 0.1–0.4% beta-DDM
Na,K-ATPase was incubated with a buffer, 150 mM NaCl, 1 mM , 4 mM NaF, 4 mM ADP, 3 mM , 2 mM glutathione, and 20 mM MOPS/n-methyl-D-glucamine (NMDG), pH 7.1, and treated with 1.95% (w/v) octaethylene glycol mono-n-dodecyl ether (C12E8) at a mass ratio (C12E8/protein) of 1.3 and separated from the insoluble fraction by centrifugation at 200.000 g and 10 uC; 17.5% PEG 2000 mme, 10% glycerol, 200 mM NaCl, 50 mM MES-NMDG
Na,K-ATPase was incubated with a buffer, 150 mM NaCl, 1 mM , 4 mM NaF, 4 mM ADP, 3 mM , 2 mM glutathione, and 20 mM MOPS/n-methyl-D-glucamine (NMDG), pH 7.1, with 0.25 mM oligomycin A and treated with 1.95% (w/v) octaethylene glycol mono-n-dodecyl ether (C12E8) at a mass ratio (C12E8/protein) of 1.3 and separated from the insoluble fraction by centrifugation at 200.000 g and 10 uC; 17.5% PEG 2000 mme, 10% glycerol, 200 mM NaCl, 50 mM MES-NMDG
16-17% PEG 2000 mme, 10% glycerol, 200 mM , 100 mM MES-NMDG, pH 6.2, 100 mM urea, 5% tert-butanol, 5 mM DTT, and 1.6 mM sucrose monodecanoate
