Table 1.
Rapamycin treatment in MKN45 Cells. MKN45 cells were SILAC labeled (light = DMSO, heavy = rapamycin; K + 8 and R + 10) and treated with either DMSO or 1mM rapamycin for 2 h. A multiplexed Affi-BAMS assay was conducted for total and phospho-specific targets. The calculated mass of the light and observed light and heavy SILAC pairs are annotated in the table along with the heavy incorporated residues (blue) and phosphorylated residues (red). The average ratio (H:L) and coefficient of variation (C.V.) is provided for each Affi-BAMS assay.
| Accession | Protein | Site/Total | Average Ratio (H:L) | C.C. (%) | Peptide | Calc. Light m/z | Light m/z | Heavy m/z |
|---|---|---|---|---|---|---|---|---|
| Q13541 | 4EBP1 | Total | 1.22 | 2.72 | K.RAGGEESQFEMDI.- | 1468.637 | 1468.83 | 1478.86 |
| P42345 | mTOR | p(S2448) | 0.15 | 5.98 | R.TRTDsYSAGQSVEILDGVELGEPAHK.T | 2839.315 | 2839.25 | 2856.99 |
| p35222 | CTNNB1 | p(S675) | 1.73 | 3.53 | K.RLsVELTSSLFR.T | 1487.762 | 1487.21 | 1507.40 |
| P31749 | AKT1 | Total | 2.89 | 11.07 | R.RPHFPQFSYSASGTA.- | 1652.782 | 1652.58 | 1662.34 |
| Q13541 | 4EBP1 | p(T37 or T46) | 3.06 | 9.09 | R.VVLGDGVQLPPGDYSTtPGGTLFSTTtPGGTR.I | 3283.599 | 3282.50 | 3302.57 |
| Q13541 | 4EBP1 | p(T37 or T46) | 0.25 | 8.57 | R.RVVLGDGVQLPPGDYSTtPGGTLFSTtPGGTR.I | 3363.566 | 3363.18 | 3383.25 |
| Q13541 | 4EBP1 | p(T65 or T70) | 0.01 | 7.61 | R.NsPVTKtPPRDLPTIPGVTSPSSDEPPMEASQSHLR.N | 3985.840 | 3985.74 | 4003.29 |
| Q92934 | BAD | p(S75) | 0.33 | 10.67 | R.HSsYPAGTEDDEGMGEEPSPFR.G | 2474.944 | 2474.96 | 2484.94 |
| P62753 | RPS6 | p(S235 & S236) | 0.04 | 6.96 | R.RLssLRASTSKSESSQK.- | 2091.887 | 2091.57 | 2128.3 |