Table I

Quantitative time course tyrosine phosphorylation profiles

A subset of protein phosphorylation sites identified from HMEC cells treated with EGF for varying duration. Results are presented from analysis of two different cell cultures; for the second cell culture, analytical replicates were also performed. The average value is presented in the table. For each protein, specific phosphorylation site(s) were identified and quantified by comparing relative peak areas of the iTRAQ marker ions. The 5-min sample was used to normalize the phosphorylation profiles. The data are therefore presented as a ratio relative to the 5-min time point. Statistical significance was calculated using a paired, two-tailed Student’s test.

IDQuantification: Sample 1, analysis 1Sample 2, average of analysis 1 and 2
ProteinSiteRatio 0 min:5 minRatio 10 min:5 minRatio 30 min:5 minRatio 0 min:5 minRatio 10 min:5 minRatio 30 min:5 min
c-CblTyr-5520.000 ± 0.001***0.916 ± 0.0150.365 ± 0.005**0.028 ± 0.066***0.715 ± 0.018***0.314 ± 0.016***
Caveolin 1Tyr-140.391 ± 0.0741.034 ± 0.0090.792 ± 0.0370.425 ± 0.027***1.121 ± 0.1360.804 ± 0.061***
EGFRTyr-11720.050 ± 0.031*0.900 ± 0.0190.505 ± 0.012*0.039 ± 0.021***0.945 ± 0.1090.662 ± 0.035***
EGFRTyr-10920.0590.7440.4290.034 ± 0.016***0.689 ± 0.111*0.461 ± 0.095**
EphB1Tyr-6000.656 ± 0.0901.142 ± 0.0451.064 ± 0.0490.733 ± 0.0381.070 ± 0.0091.182 ± 0.003**
EphrinB2Tyr-3040.8371.3641.1560.792 ± 0.1530.902 ± 0.2341.072 ± 0.190
EPS15Tyr-8490.0801.4090.6520.043 ± 0.050*1.412 ± 0.1300.605 ± 0.046
FAKTyr-5760.637 ± 0.051***1.424 ± 0.051***1.629 ± 0.036***0.569 ± 0.088***0.801 ± 0.076***1.678 ± 0.206***
FRKTyr-4970.6141.2411.3400.495 ± 0.050*1.349 ± 0.0530.773 ± 0.037
GIT 1Tyr-5450.575 ± 0.0571.183 ± 0.016*1.093 ± 0.0400.807 ± 0.063*1.212 ± 0.2761.074 ± 0.326
GSK-3-βTyr-2790.846 ± 0.014**1.112 ± 0.024*0.931 ± 0.0280.908 ± 0.1151.147 ± 0.1220.980 ± 0.082
HER2Tyr-12480.1410.7320.5320.2140.8080.492
p130CasTyr-3271.1261.1361.4422.116 ± 0.130**1.418 ± 0.058**1.497 ± 0.045**
ACK1Tyr-8570.1330.9970.3290.099 ± 0.037*0.561 ± 0.0690.358 ± 0.109
PaxillinTyr-1180.528 ± 0.1310.755 ± 0.1370.792 ± 0.1640.726 ± 0.045***0.732 ± 0.105**0.850 ± 0.095*
Plakophilin 3Tyr-840.1940.7780.1440.038 ± 0.007**0.429 ± 0.015*0.286 ± 0.023*
PLC-γTyr-775, Tyr-7830.0430.6050.3350.008 ± 0.016**0.287 ± 0.013**0.231 ± 0.001***
PTP NR 11Tyr-620.945 ± 0.0321.266 ± 0.023*1.122 ± 0.0161.074 ± 0.1570.971 ± 0.1421.128 ± 0.202
PTPR ATyr-7980.833 ± 0.005*0.943 ± 0.0100.708 ± 0.003**0.873 ± 0.073*0.740 ± 0.119*0.613 ± 0.103**
Rho GEF 5Tyr-19, Tyr-220.019 ± 0.037*0.844 ± 0.0520.632 ± 0.0470.000 ± 0.012**0.352 ± 0.012**0.248 ± 0.040*
SHCTyr-239, 2400.147 ± 0.015**1.160 ± 0.0830.902 ± 0.0590.173 ± 0.023*0.995 ± 0.1740.734 ± 0.198
SHCTyr-3180.028 ± 0.046*1.077 ± 0.0260.646 ± 0.003**0.296 ± 0.3920.878 ± 0.2170.908 ± 0.543
SHIP-2Tyr-9860.043 ± 0.054***0.801 ± 0.0800.363 ± 0.028**0.013 ± 0.045***0.428 ± 0.160**0.291 ± 0.160**
STAM-1Tyr-3810.3092.6590.9260.148 ± 0.288*4.188 ± 0.724**1.624 ± 0.397
STAM-2Tyr-3740.2182.7530.9080.459 ± 0.4742.515 ± 0.613*1.308 ± 0.017***
STAT-3 Isoform 1Tyr-7050.209 ± 0.026***0.803 ± 0.073*0.244 ± 0.046***0.242 ± 0.092**0.565 ± .039**0.478 ± 0.134*
STAT-3 Isofrom2Tyr-7040.1950.7940.2750.196 ± 0.1970.545 ± 0.1780.406 ± 0.266
  • * , p < 0.05;

  • ** , p < 0.01;

  • *** , p < 0.001.