Figure 5.

Effect of kinase activity and amino acids 353–835 in the C-terminal region of GLK on JNK activation. (A) 293 cells were cotransfected with vector alone (lane 1), wild-type GLK (lane 2), K35E mutant (lane 3), or a GLK C-terminal deletion mutant (ΔGLK) (lane 4) plus JNK. JNK activity was assayed using GST c-Jun as substrate. (B) Lysates containing equal amounts of total protein were run on 10% SDS/PAGE and transferred to a nitrocellulose membrane. The blot was probed with a mAb against the flag epitope followed by rabbit anti-mouse IgG. The blot was developed using the ECL system. (C) Kinase activity of GLK C-terminal deletion mutant. Lysates from cells transfected with vector alone (lane 1) or with GLK C-terminal deletion mutant (ΔGLK) (lane 2) were prepared and recombinant protein was immunoprecipitated with mAb M2 followed by kinase assay with MBP as substrate.