Proteomic analysis of differential protein expression in Sod2+/+, Sod2-/-, and antioxidant-treated Sod2-/- RBC membrane preparations
Protein ID . | SOD2+/+ versus SOD2-/- . | SOD2-/- (EUK-189) versus SOD2-/- . |
|---|---|---|
| Membrane/structural proteins | ||
| Anion exchanger 4b | -5.95 | -1.75 |
| Guanine nucleotide exchange factor | -19.17 | -1.03 |
| p97 membrane-bound transferrin-like protein | -54.20 | -1.01 |
| Actin-like protein | — | 1.96 |
| Rbcc 728 | -5.63 | 1.32 |
| Alpha2-macroglobulin receptor | — | 8.36 |
| HA receptor for hyaluronic acid | -25.62 | 1.41 |
| HA receptor for hyaluronic acid* | -18.59 | 1.99 |
| Surfeit locus protein 4 | — | 2.26 |
| Cytokeratin 10 | 3.79 | 17.16 |
| Kainate receptor GluR7 subunit | — | -1.50 |
| Stress response/chaperones | ||
| 60-kDa chaperonin mitochondrial | — | -2.40 |
| Heat-shock 60-kDa chaperonin | — | -1.93 |
| Sulfotransferase-related protein | — | 1.35 |
| Prohibitin | — | -1.82 |
| Prohibitin* | — | -1.56 |
| Peroxiredoxin 2 | 1.48 | 2.90 |
| Peroxiredoxin 2* | — | 2.75 |
| Peroxiredoxin 2* | — | 2.79 |
| DNAK-type molecular chaperone | -10.11 | -3.15 |
| Metabolism | ||
| Octanoyltransferase | — | 2.05 |
| Gamma enolase | -23.01 | 1.90 |
| Gamma enolase* | — | 1.67 |
| Microsomal tryglyceride transfer protein | — | 4.25 |
| Microsomal tryglyceride transfer protein* | -5.12 | 3.50 |
| UDP glucuronyltransferase-P | — | 2.06 |
| Carbonic anhydrase I | 1.55 | 5.54 |
| (Similar to) biliverdinreductase B flavin reductase | — | 2.67 |
| Leucyl-tRNA synthetase | -4.66 | -1.00 |
| Nucleoside diphosphate kinase | — | 2.68 |
| Mitochondrial/bioenergetics | ||
| GFAP | — | 1.38 |
| MECA32 | — | 1.39 |
| Delta subunit ATPase | — | -2.17 |
| Cytochrome C oxidase, subunit Va | — | -2.52 |
| ATP synthase beta subunit | -1.60 | -1.95 |
| ATP synthase Beta chain | — | 4.01 |
| ATP synthase D chain | -24.02 | -2.03 |
| ATP synthase D chain* | -1.96 | -1.32 |
| ATP synthase alpha, mitochondrial | — | -4.67 |
| Voltage-dependent anion channel† | 1.00 | 1.00 |
| Not classified | ||
| Calreticulin | — | 1.46 |
| Cleavage and polyadenylation-specific factor 2 | -2.96 | 2.43 |
| Thrombospondin | -8.87 | 1.36 |
| Thrombospondin* | -5.34 | 1.25 |
| Cytochrome B5 | — | -1.11 |
| Apolipoprotein A-I | — | 2.44 |
| Retinoic acid-binding protein II | — | 2.44 |
| Ubiquitin B | -1.02 | 1.17 |
| Trophoblast protein-1 | -1.34 | -1.98 |
Protein ID . | SOD2+/+ versus SOD2-/- . | SOD2-/- (EUK-189) versus SOD2-/- . |
|---|---|---|
| Membrane/structural proteins | ||
| Anion exchanger 4b | -5.95 | -1.75 |
| Guanine nucleotide exchange factor | -19.17 | -1.03 |
| p97 membrane-bound transferrin-like protein | -54.20 | -1.01 |
| Actin-like protein | — | 1.96 |
| Rbcc 728 | -5.63 | 1.32 |
| Alpha2-macroglobulin receptor | — | 8.36 |
| HA receptor for hyaluronic acid | -25.62 | 1.41 |
| HA receptor for hyaluronic acid* | -18.59 | 1.99 |
| Surfeit locus protein 4 | — | 2.26 |
| Cytokeratin 10 | 3.79 | 17.16 |
| Kainate receptor GluR7 subunit | — | -1.50 |
| Stress response/chaperones | ||
| 60-kDa chaperonin mitochondrial | — | -2.40 |
| Heat-shock 60-kDa chaperonin | — | -1.93 |
| Sulfotransferase-related protein | — | 1.35 |
| Prohibitin | — | -1.82 |
| Prohibitin* | — | -1.56 |
| Peroxiredoxin 2 | 1.48 | 2.90 |
| Peroxiredoxin 2* | — | 2.75 |
| Peroxiredoxin 2* | — | 2.79 |
| DNAK-type molecular chaperone | -10.11 | -3.15 |
| Metabolism | ||
| Octanoyltransferase | — | 2.05 |
| Gamma enolase | -23.01 | 1.90 |
| Gamma enolase* | — | 1.67 |
| Microsomal tryglyceride transfer protein | — | 4.25 |
| Microsomal tryglyceride transfer protein* | -5.12 | 3.50 |
| UDP glucuronyltransferase-P | — | 2.06 |
| Carbonic anhydrase I | 1.55 | 5.54 |
| (Similar to) biliverdinreductase B flavin reductase | — | 2.67 |
| Leucyl-tRNA synthetase | -4.66 | -1.00 |
| Nucleoside diphosphate kinase | — | 2.68 |
| Mitochondrial/bioenergetics | ||
| GFAP | — | 1.38 |
| MECA32 | — | 1.39 |
| Delta subunit ATPase | — | -2.17 |
| Cytochrome C oxidase, subunit Va | — | -2.52 |
| ATP synthase beta subunit | -1.60 | -1.95 |
| ATP synthase Beta chain | — | 4.01 |
| ATP synthase D chain | -24.02 | -2.03 |
| ATP synthase D chain* | -1.96 | -1.32 |
| ATP synthase alpha, mitochondrial | — | -4.67 |
| Voltage-dependent anion channel† | 1.00 | 1.00 |
| Not classified | ||
| Calreticulin | — | 1.46 |
| Cleavage and polyadenylation-specific factor 2 | -2.96 | 2.43 |
| Thrombospondin | -8.87 | 1.36 |
| Thrombospondin* | -5.34 | 1.25 |
| Cytochrome B5 | — | -1.11 |
| Apolipoprotein A-I | — | 2.44 |
| Retinoic acid-binding protein II | — | 2.44 |
| Ubiquitin B | -1.02 | 1.17 |
| Trophoblast protein-1 | -1.34 | -1.98 |
Table 2 lists 41 proteins identified by MALDI-MS as differentially expressed when comparing membrane preparations from Sod2+/+, Sod2-/-, and antioxidant-treated Sod2-/- RBCs. Sod2+/+ mice were made anemic through serial phlebotomy in order to ensure that reticulocyte counts were comparable to those in Sod2-/- samples. Data have been normalized to content of the mitochondrial protein voltage-dependent anion channel (VDAC), in bold. Most of the identified proteins are expressed at higher levels in Sod2-/- RBCs than in Sod2+/+ RBCs (negative values in the “SOD2+/+ versus SOD2-/-” column indicate fold decrease in protein expression). Changes in protein expression in response to Euk-189 therapy vary (“SOD2-/- (EUK-189) versus SOD2-/-” column). Some proteins were not detected in the Sod2+/+ sample, and data for these proteins is limited to comparison of plus and minus Euk-189 treatment. — indicates protein not detected in Sod2+/+ preparation.
Several proteins migrated as 2 or more unique spots, suggesting differences in posttranslational modification or variant splicing.
This row used for normalization.