Table II . Relative amounts of FLNa and Talin 1 based on chromatographic peak areas of five selected specific peptides.
PLB985/MT-FLAG-ASB2wt and PLB985/MT-FLAG-ASB2LA cells were treated with 100 μm ZnSO4 for 8 h. Cytosolic cell extracts of two independent experiments were separated by SDS-PAGE. Filamin A and Talin 1 were identified by shotgun proteomics analysis, and their relative quantification was performed with the five specific peptides of each protein described in supplemental Data 7 and 8. For each peptide ion of FLNa and Talin 1, XICs were extracted from four MS analyses. Ratios indicated for each biological and technical replicate correspond to ratios of the sum of the chromatographic peak areas measured in slices where the protein was identified in ASB2LA-expressing cell extracts to the sum of the chromatographic peak area measured in slices where the protein was identified in ASB2wt-expressing cell extracts. a, b, c, and d are technical replicates from two biological replicates. Results are mean and S.D. from two independent experiments.
| Peptide | Experiment |
Average | |||
|---|---|---|---|---|---|
| a | b | c | d | ||
| FLNa | |||||
| EAGAGGLAIAVEGPSK | 4.6 | 8.4 | 8.0 | 8.0 | |
| IANLQTDLSDGLR | 4.8 | 9.5 | 9.2 | 8.9 | |
| VANPSGNLTETYVQDR | 8.2 | 8.8 | 12.1 | 12.5 | |
| AFGPGLQGGSAGSPAR | 7.1 | 11.2 | 11.9 | 11.9 | |
| VTAQGPGLEPSGNIANK | 6.6 | 10.1 | 9.3 | 9.3 | |
| 6.3 ± 1.5a | 9.6 ± 1.1a | 10.1 ± 1.8a | 10.1 ± 2.0a | 9.0 ± 1.9 | |
| Talin 1 | |||||
| LGAASLGAEDPETQVVLINAVK | 1.1 | 0.7 | 0.9 | 1.0 | |
| ERIPEAPAGPPSDFGLFLSDDDPK | 1.2 | 2.1 | 1.0 | 1.2 | |
| EADESLNFEEQILEAAK | 0.9 | 0.7 | 0.9 | 1.0 | |
| ILAQATSDLVNAIK | 0.5 | 1.0 | 0.8 | 1.3 | |
| LAQAAQSSVATITR | 0.6 | 0.8 | 0.9 | 0.9 | |
| 0.8 ± 0.3a | 1.1 ± 0.6a | 0.9 ± 0.1a | 1.1 ± 0.1a | 1.0 ± 0.2 | |
a Means and S.D. of the five different peptides of FLNa and Talin 1 in experiments a–d.