Proteins affected by treatment with DNR
| Spotnumber . | Proteinabbreviation . | pI . | Molecular weight . | Relative spot intensity . | |||
|---|---|---|---|---|---|---|---|
| Observed . | Predicted . | Observed . | Predicted . | Control, in cellulo/in vitro . | DNR, in cellulo/in vitro . | ||
| 1 | rpP2 | 4.48/4.39/4.3 | 4.42/4.37/4.31 | 13 | 12 | 5.0 | 26 |
| 2 | PDI | 4.75 | 4.76 | 58 | 57 | >5.0 | >20 |
| 3 | PCNA | 4.57 | 4.57 | 31 | 29 | 4.4 | 12 |
| 4 | cutA | 4.45 | 5.15 | 15 | 17 | n.d. | n.d. |
| 5 | CALR | 4.31 | 4.29 | 46 | 48 | 1.5 | 2.0 |
| 6 | EF1Ba | 4.50 | 4.50 | 25 | 25 | 0.89 | 1.8* |
| 7 | PSMA5 | 4.63 | 4.74 | 24 | 26 | 0.21 | 0.62 |
| 8 | CTM | 4.65 | 4.80 | 33 | 28 | 0.98 | 0.36 |
| 9 | MLC | 4.32 | 4.46 | 16 | 17 | 0.46 | 0.25 |
| 10 | n.i. | 4.62 | n.d. | 20 | n.d. | 0.12 | 0.11 |
| 11 | n.i. | 4.73 | n.d. | 15 | n.d. | 0.09 | 0.03 |
| Spotnumber . | Proteinabbreviation . | pI . | Molecular weight . | Relative spot intensity . | |||
|---|---|---|---|---|---|---|---|
| Observed . | Predicted . | Observed . | Predicted . | Control, in cellulo/in vitro . | DNR, in cellulo/in vitro . | ||
| 1 | rpP2 | 4.48/4.39/4.3 | 4.42/4.37/4.31 | 13 | 12 | 5.0 | 26 |
| 2 | PDI | 4.75 | 4.76 | 58 | 57 | >5.0 | >20 |
| 3 | PCNA | 4.57 | 4.57 | 31 | 29 | 4.4 | 12 |
| 4 | cutA | 4.45 | 5.15 | 15 | 17 | n.d. | n.d. |
| 5 | CALR | 4.31 | 4.29 | 46 | 48 | 1.5 | 2.0 |
| 6 | EF1Ba | 4.50 | 4.50 | 25 | 25 | 0.89 | 1.8* |
| 7 | PSMA5 | 4.63 | 4.74 | 24 | 26 | 0.21 | 0.62 |
| 8 | CTM | 4.65 | 4.80 | 33 | 28 | 0.98 | 0.36 |
| 9 | MLC | 4.32 | 4.46 | 16 | 17 | 0.46 | 0.25 |
| 10 | n.i. | 4.62 | n.d. | 20 | n.d. | 0.12 | 0.11 |
| 11 | n.i. | 4.73 | n.d. | 15 | n.d. | 0.09 | 0.03 |
Cells were treated with vehicle or 8 μmol/L DNR for 4.5 hours, pulse-labeled for the last 30 minutes with [35S]methionine, and protein spots 1 to 11 in 2DE gels (see Figure 4 for details) analyzed for relative labeling intensity. Proteins translated from mRNA isolated from vehicle-treated (Ctr) or DNR-treated cells were similarly separated and analyzed. The ratio between relative spot intensity in cells (in cellulo) and after in vitro translation (in vitro) is listed in the two rightmost columns. The data are averages from 3 separate experiments. Protein abbreviations refer to the following proteins (Swissprot accession numbers in parentheses). rpP2 indicates 60S acidic ribosomal protein P2 (P05387); PDI, protein disulfide isomerase/prolyl 4-hydroxylase β subunit (NP_000909); PCNA, proliferating cell nuclear antigen (NP_002583); cutA, divalent cation tolerance homolog isoform 2(NP_057005); CALR, calreticulin precursor (NP_004334); eEF1Ba, elongation factor 1Ba (P24534); PSMA5, proteasome endopeptidase complex, ζ chain (S17521); CTM, cytoskeletal tropomyosin (CAA28258); MLC, myosin light polypeptide 6 (P60662); n.d., not determined; and n.i., not identified.
The value for EF1Ba is slightly underestimated due to cleavage of this protein (see Figure 6).