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. 2012 Dec 20;7(12):e51817. doi: 10.1371/journal.pone.0051817

Table 2. Quantification of HER2 phosphorylation and the differences after treatment.

Site* % Phosph. Difference From Basal
Trastuzumab Bis-Fab 1325 Heregulin
T701 48.2 −0.4 18.1 20.3
S728 0.6 0.1 0 0
Y735 0.1 0 0 0
Y877 0.1 0 0 0
Y1005 29.6 −6.8 15.3 18.8
S1054 87.7 1.1 3.7 4.2
S1073 29.3 −5.8 17.1 15.1
S1078 35.9 6.8 9.6 10.4
S1083 37.1 9.3 6.3 7.5
S1100 1.3 −0.3 0.1 0.4
T1103 1.3 0.5 0.1 0.4
Y1139 84.1 2.8 10.3 6.7
Y1151 6.8 3.9 5.4 −1.3
T1240 40.6 −15.1 −22.3 −9.8
Y1248 17.8 21.1 14.9 −4
*

Phosphorylation sites were identified by HER2 phosphomapping.

Quantitative mass spectrometry (MS) was done to determine the basal level of phosphorylation at each site. The absolute amount of phosphorylated peptide was compared to the total of phosphorylated and non-phosphorylated peptides to calculate the percent phosphorylation (% Phosph.)

Ten minutes after treatment with the indicated molecule, quantitative MS was done to determine the % phosphorylation. This was subtracted from the % phosphorylation in the basal state to provide the mean difference after treatment reported here. This time point was chosen because maximal phosphorylation of AKT was shown to occur within ten minutes after agonist treatment ( Figure 4 ).