Mapping the interaction of TrkA with GIPC
deletion mutants. (A) Schematic representation of mouse GIPC deletion
mutants. The interaction between GIPC mutants and
TrkA448–522 shown on the right is based on data from B.
(+, interaction; −, no interaction). (B) Left, in vitro interaction of
TrkA juxtamembrane region with GIPC truncation mutants.
GIPC81–333 (lane 2), GIPC125–333(PDZ+C) (lane
4), and GIPC125–225(PDZ) (lane 10) bind to
GST-TrkA448–522 but not to GST alone (lanes 1, 3, and 9),
whereas GIPC1–124(N) (lane 6) or
GIPC226–333(C) (lane 8) did not bind to
GST-TrkA448–522. GST-TrkA448–522 or GST alone
immobilized on glutathione-Sepharose beads was incubated with the
indicated in vitro translated, 35S-labeled GIPC mutant
proteins. The 35S-labeled proteins bound to the beads were
separated by SDS-PAGE and detected by autoradiography. Right, an
aliquot of each in vitro translated product was resolved by SDS-PAGE
and examined by autoradiography to confirm the protein yield and the
molecular size.