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Figure 2.
Fig. 2. Small NH[2]-terminal region of JIP-1 is sufficient
for interaction with JNK. (A) Cell lysates containing
Flag-epitope-tagged^ JNK1 were incubated with GST and GST-JIP-1
(residues 127 to 281) bound to glutathione-Sepharose, and bound
proteins were detected^ by protein immunoblot analysis with an
antibody to Flag. The effect of increasing concentrations (0, 4,
8, 16, 32, and 64 µg/ml) of^ synthetic peptide
corresponding to JIP-1 residues 148 to 174 or to a peptide with
a scrambled sequence (control) was examined. (B) Binding of JNK1
to GST and GST-JIP-1 (residues 127^ to 281) was examined in the
absence and presence of synthetic^ peptides (64 µg/ml).
The effect of the wild-type peptide (JIP-1^ residues 148 to
174), peptides with point mutations (substitution with Gly)
[Thr159  Gly159
(T159G)] (18), and a peptide with a scrambled primary sequence^
(control) was examined. The mutant JIP(159-162G) was
constructed^ by replacement of JIP-1 residues 159 to 162 with
Gly.
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