Table 2.
HPLC-ESI-MS/MS analysis of endogenous IRS-1 interaction partners in L6 myotubes upon insulin stimulation. Data shown are mean ± SEM, n=5
| (A) Spectrum count (SC) of endogenous IRS-1 interaction partners in the basal (BAS) and insulin (INS) stimulated samples | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Gene symbol a | Molecular weight (Da) | SC BAS | SC INS | Exp.1 BAS | Exp.1 INS | Exp.2 BAS | Exp.2 INS | Exp.3 BAS | Exp.3 INS | Exp.4 BAS | Exp.4 INS | Exp.5 BAS | Exp.5 INS |
| ACTN1* | 102946 | 10±4 | 22±6 | 13 | 34 | 20 | 28 | 15 | 32 | 0 | 6 | 0 | 10 |
| ACTN4* | 104900 | 20±8 | 35±7 | 21 | 49 | 34 | 44 | 38 | 46 | 4 | 14 | 1 | 20 |
| CALD1* | 60567 | 11±4 | 21±6 | 23 | 41 | 14 | 27 | 17 | 22 | 1 | 13 | 1 | 3 |
| EEF2* | 95268 | 6±3 | 11±3 | 4 | 9 | 12 | 16 | 13 | 17 | 0 | 5 | 1 | 6 |
| HSP90AB1* | 83266 | 16±4 | 27±4 | 16 | 25 | 29 | 32 | 14 | 23 | 4 | 17 | 16 | 38 |
| HSPA5* | 72331 | 30±5 | 44±5 | 34 | 52 | 36 | 42 | 41 | 59 | 12 | 33 | 25 | 32 |
| HTRA1* | 51312 | 4±2 | 10±2 | 4 | 8 | 10 | 14 | 4 | 12 | 1 | 2 | 1 | 15 |
| MPRIP* | 257845 | 53±18 | 92±10 | 71 | 95 | 100 | 105 | 73 | 117 | 11 | 58 | 11 | 83 |
| PABPC1* | 70683 | 3±1 | 5±1 | 4 | 5 | 4 | 6 | 6 | 9 | 1 | 1 | 1 | 4 |
| PPP1R12A* | 115268 | 5±2 | 12±1 | 5 | 12 | 12 | 13 | 7 | 16 | 1 | 9 | 0 | 9 |
| TNNT3* | 30734 | 17±7 | 25±6 | 18 | 22 | 25 | 28 | 38 | 46 | 0 | 21 | 2 | 6 |
| (B) Normalized spectrum count (NormSC) of endogenous IRS-1 interaction partners in L6 myotubes from basal (BAS) and insulin (INS) stimulated samples | |||||||||||||
| Gene Symbol a | NormSC BAS | NormSC INS | Exp.1 BAS | Exp.1 INS | Exp.2 BAS | Exp.2 INS | Exp.3 BAS | Exp.3 INS | Exp.4 BAS | Exp.4 INS | Exp.5 BAS | Exp.5 INS | |
| ACTN1* | 0.104±0.046 | 0.216±0.062 | 0.14 | 0.32 | 0.24 | 0.33 | 0.14 | 0.25 | 0.00 | 0.07 | 0.00 | 0.11 | |
| ACTN4* | 0.206±0.081 | 0.344±0.088 | 0.23 | 0.45 | 0.40 | 0.52 | 0.36 | 0.37 | 0.03 | 0.15 | 0.01 | 0.23 | |
| CALD1* | 0.12±0.048 | 0.21±0.068 | 0.25 | 0.38 | 0.17 | 0.32 | 0.16 | 0.18 | 0.01 | 0.14 | 0.01 | 0.03 | |
| EEF2* | 0.062±0.029 | 0.108±0.03 | 0.04 | 0.08 | 0.14 | 0.19 | 0.12 | 0.14 | 0.00 | 0.06 | 0.01 | 0.07 | |
| HSP90AB1* | 0.166±0.05 | 0.282±0.069 | 0.17 | 0.23 | 0.34 | 0.38 | 0.13 | 0.18 | 0.03 | 0.19 | 0.16 | 0.43 | |
| HSPA5* | 0.302±0.063 | 0.432±0.083 | 0.37 | 0.48 | 0.42 | 0.49 | 0.39 | 0.47 | 0.08 | 0.36 | 0.25 | 0.36 | |
| HTRA1* | 0.044±0.02 | 0.106±0.032 | 0.04 | 0.07 | 0.12 | 0.17 | 0.04 | 0.10 | 0.01 | 0.02 | 0.01 | 0.17 | |
| MPRIP* | 0.566±0.209 | 0.926±0.189 | 0.76 | 0.88 | 1.18 | 1.24 | 0.70 | 0.93 | 0.08 | 0.64 | 0.11 | 0.94 | |
| PABPC1* | 0.034±0.01 | 0.05±0.013 | 0.04 | 0.05 | 0.05 | 0.07 | 0.06 | 0.07 | 0.01 | 0.01 | 0.01 | 0.05 | |
| PPP1R12A* | 0.054±0.025 | 0.118±0.023 | 0.05 | 0.11 | 0.14 | 0.15 | 0.07 | 0.13 | 0.01 | 0.10 | 0.00 | 0.10 | |
| TNNT3* | 0.172±0.072 | 0.24±0.064 | 0.19 | 0.20 | 0.29 | 0.33 | 0.36 | 0.37 | 0.00 | 0.23 | 0.02 | 0.07 | |
ACTN1 = ↦-actinin-1; ACTN4 = ↦-actinin-4; CALD1 = caldesmon; CENTG2 = centaurin γ 2; EEF2 = elongation factor 2; FLII = flightless I homolog; HSP90AB1 = heat shock protein HSP 90-β; HSPA5 = 78 kDa glucose-regulated protein; HTRA1 = HtrA serine peptidase 1 isoform CRA a; LRRFIP2 = leucine-rich repeat flightless-interacting protein 2; MPRIP = myosin phosphatase Rho-interacting protein; PABPC1 = polyadenylate-binding protein 1; PPP1R12A = protein phosphatase 1 regulatory subunit 12A; TNNT3 = isoform 1 of troponin T, fast skeletal muscle
Proteins have five increases out of the five comparisons
P<0.05 based on the sign test. There were eleven such proteins