Table 3.
Quantitative analysis of differentially expressed protein spots by 2DE using pH3-10 range (this study) and pH4-7 platform (previous work, ref.[17])
| No.1 | Ref.2 | Name3 | IPG 4-74 |
IPG 3-105 |
Nephge 3-106 |
||
|---|---|---|---|---|---|---|---|
| Standard5 | High load5 | Standard6 | High load6 | ||||
| a |
1 |
SSA1/2 |
2.4 ± 0.2 |
1,6 ± 0,1 |
1,6 ± 0,4 |
2,6 ± 0,3 |
2,0 ± 0,2 |
| b |
2 |
SSA1/2 |
|||||
| c |
3 |
SSA4 |
|||||
| d |
4 |
KAR2 |
3.8 ± 0.4 |
2,7 ± 0,5 |
1,8 ± 0,4 |
9,0 ± 3,1 |
2,5 ± 0,2 |
| e |
5 |
SSE1 |
2.3 ± 0.2 |
1,4 ± 0,3 |
1,8 ± 0,7 |
2,2 ± 0,8 |
1,7 ± 0,1 |
| f |
6 |
HSC82 |
2.1 ± 0.3 |
2,0 ± 0,2 |
2,1 ± 1,0 |
- - |
- - |
| 6 |
HSP82 |
||||||
| g |
7 |
ENO2 |
1.5 ± 0.2 |
1,4 ± 0,3 |
1,1 ± 0,2 |
1,3 ± 0,2 |
1,1 ± 0,1 |
| h |
N.I.7 |
SSA1/27 |
2.2 ± 0.3 |
1,5 ± 0,1 |
1,1 ± 0,1 |
1,6 ± 0,3 |
2,1 ± 0,4 |
| ?8 |
N.A. |
GPM1 |
N.A. |
2,2 ± 1,3 |
0,7 ± 0,2 |
1,3 ± 0,3 |
1,0 ± 0,1 |
| !8 | N.A. | SIS1 | N.A. | - - | - - | 2,6 ± 0,4 | 2,2 ± 0,2 |
1Differentially expressed protein spots in this experiment are indicated by letters (see Figures 1 and 2).
2The same protein spots are indicated by the numbers in the referenced article ([17], see Figure nine and Table one).
3Accepted name from the Saccharomyces genome database (SGD) and YPD. Spots 1 and 2 represent mixtures of similar proteins Ssa1 and Ssa2 (97% identity) at an unknown ratio (see Table one legend in [17]).
4Cellular protein expression fold change in MeH expressing versus control cells that was determined in previous work using pH4-7 IPG-based 2DE system (Invitrogen); the values are taken from the Table one in reference [17].
5Expression fold changes of the same proteins determined from independent experiments in this work using pH3-10 IPG strips (Invitrogen); ~50 μg of whole cell protein was loaded onto IPG strips in “Standard” experiment, whereas ~100 μg was used in a “High load” experiment.
6Expression fold changes of the same proteins determined from independent experiments in this work using pH3-10 NEPHGE first dimension gels (WITAvision); ~30 μg of whole cell protein was loaded onto NEPHGE gels in “Standard” experiment, whereas ~100 μg was used for each gel in the “High load” analysis.
7Not identified (N.I.) in previous study, because increased amount of this protein was observed only in cells expressing MeH, but not MuHN protein (the expression fold change in MeH/Control cells determined by IPG4-7 system here is given from our unpublished data).
8? and ! indicate basic protein spots (pI > 7) that were not analysed in previous experiment on pH4-7 platform (N.A. – not assayed). Despite that protein spot “?” showed false expression change (“artefact”) in IPG-based system (unreliable expression changes are apparent by high error range), in this experiment it was identified as phosphoglycerate mutase 1 (Gpm1p). Protein Sis1p in this study was identified using NEPHGE-based 2DE system, whereas it was not detected by IPG-based 2DE method.