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Table 1 Kinase-substrate pairs.

From: Protein kinase substrate identification on functional protein arrays

   Solution Assay Protein Array Assay
    Z_Score
Upstream Kinases Substrates (Signal/Background) -BSA +BSA
CaMKII Tau Protein 16.01 15.00 10.1
CDK5/p35 Tau Protein 28.24 18.80 6.5
CK2 pTEN 67.65 12.80 18.6
ERK2(MAPK) 4EBP/PHAS-1 3.85 6.10 9.6
ERK2(MAPK) Elk-1 Fusion Protein 8.17 17.00 20
ERK2(MAPK) Tau Protein 3.34 24.10 19.4
GSK3beta GST-Axin 33.63 1.40 2.4
JNK1 c-Jun 1–79 6.84 19.20 103.3
MAP2K6(MKK6) MAPK12 Inactive 20.45 0.90 0.1
MAP2K6(MKK6) MAPK14, Inactive 42.02 12.50 23.6
MAPK14 (p38alpha) 4EBP/PHAS-1 3.58 9.90 17.5
MAPK14 (p38alpha) ATF2 (aa 19–96) 6.48 22.60 25.5
MAPK14 (p38alpha) MAPKAP-K2 Inactive 13 13.90 19.4
MAPK14 (p38alpha) MAPKAP-K3 Inactive 6.21 10.70 22.4
MAPK14 (p38alpha) MAPKAP-K5, Inactive 5.07 5.40 2.4
MKK4/SKK1 MAPK12 Inactive 17.43 1.80 7.7
MKK4/SKK1 MAPK8 (JNK1) Inactive 23.17 2.90 0.7
MKK4/SKK1 MAPK9 (JNK2) Inactive 6.61 0.90 5.9
p38β2/SAPK2b2 MAPKAP-K3 Inactive 12.63 3.40 9.4
PDK1 AKT2 Inactive 6.33 4.30 3.1
PKA ATF2 (aa 19–96) 8.34 3.40 0.7
PKA Tau Protein 12.66 33.90 4.6
ROKα/ROCK-II MYPT1 (654–880) 51.54 14.40 23
RPS6KA3(RSK2) Estrogen Receptor-α 9.08 0.40 2.7
  1. A set of 24 kinase-substrate interactions comprising 14 different protein kinases and 18 substrates was defined as the test set for benchmarking protein microarray results against solution phase assays. Solution phase reactions were run for each of these pairs in the presence of 33P-ATP. After 1 hour, reactions were terminated by the addition of sample buffer and run on SDS-PAGE gels. Gels were subsequently dried, exposed to a phosphoscreen, and imaged in a Cyclone phosphoimager. Pixel intensity data from the resultant high resolution images was extracted and used to calculate the reported signal/background values. Protein microarray assays were run by incubating arrays with exogenous kinase in the presence of 33P-ATP. After 60 minutes, arrays were washed, dried, exposed to a phosphoscreen, and imaged using a Cyclone phosphoimager. Pixel intensity data from the resultant high resolution images was extracted using GenePix software, and used to calculate the reported Z-Scores.
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