Table 1.

Identification and in vitro characterization of protein kinases targeted by gefitinib

Protein kinaseGi no.MW (kDa)Gel identification *LC-MS/MS identification Binding assay Kinase assay IC50 (μmol/L)
Aurora A744641145,790+++60 ± 7
Aurora B475917839,280+ND11.1 ± 2.9
BLK91420457,757+ND3.1 ± 0.4
Bub12981233122,389++ND
BRK517464751,834+++0.82 ± 0.06
CaMKII §13.5 ± 1.4
CaMKIIα2625171254,088+NDND
CaMKIIβ413926857,914+NDND
CaMKIIγ2666719958,365++NDND
CaMKIIδ2666718354,128++NDND
CK1δ751233147,374+ND61 ± 7
CK1ε450309347,315++ND
CSK475807850,704+++41 ± 17
EGFR29725609134,277+++0.014 ± 0.5 × 103
EphB4495473108,334++ND1.22 ± 0.49
Fyn450382360,762++ND
GAK4885251143,165+++0.090 ± 0.018
HCK30683257,296++2.09 ± 0.35
ILK475860651,419++ND
IKKε728887880,462+++35% at 100 μmol/L
JNK2108226648,149+++39% at 100 μmol/L
Lyn450505558,574++0.95 ± 0.46
Met3792812735,259++3.2 ± 1.1
p38α249960041,293+++1.19 ± 0.03
RICK450653761,195+++0.049 ± 0.001
Tnk1409207971,926++ND
Yes488566160,801+++1.75 ± 0.13
  • * Identified protein spots from Fig. 1 that contained protein kinases are included; for a full list of all analyzed protein spots, see Supplementary Table 1.

  • LC-MS/MS data of all identified proteins are listed in Supplementary Table 2.

  • In vitro binding assays are shown in Supplementary Fig. 1.

  • § CaMKII activity was analyzed with an enzyme preparation that contains a mixture of different isoforms.

  • Percent inhibition of kinase activity in the presence of 100 μmol/L gefitinib is shown.

  • NOTE: Abbreviations: BLK, B-lymphoid tyrosine kinase; CK1δ, casein kinase 1δ; HCK, hemopoietic cell kinase; ILK, integrin-linked kinase; ND, not determined.