Figure 6: Amino acid following NPEY motif dictates the preference of Shc binding.
From: Domain-dependent effects of insulin and IGF-1 receptors on signalling and gene expression
(a) Alignment of the juxtamembrane regions in the insulin and IGF-1 receptors throughout evolution. (b) Co-immunoprecipitation of IR, Shc and IRS-1. Flag-tagged IR or mutated IR were immunoprecipitated with anti-Flag antibody following insulin stimulation. Bound Shc and IRS-1 was detected by biotin-HA immunoblotting. Phosphorylation of proteins from total cell lysates was detected by phospho-specific antibodies. (c) Co-immunoprecipitation of IR and IRS-1 with increasing expression levels of Shc. Flag-tagged IR or mutated IR were immunoprecipitated with anti-Flag antibody following insulin stimulation. Bound Shc and IRS-1 was detected by biotin-HA immunoblotting. (d) Co-immunoprecipitation of Shc and IR. HA-tagged Shc was immunoprecipitated with anti-HA antibody following insulin stimulation. Bound IR and IRL973F were detected by anti-Flag immunoblotting. (e) Solution NMR structure of the Shc-PTB domain (green and blue) bound to a phosphorylated segment containing the NPxY motif from the TrkA receptor (brown). Zoom-in: detailed view of the interaction between Shc-PTB and Phe at TrkA Y+1 position. Note, Phe makes extensive van der Waals contacts in the hydrophobic pocket formed by Phe152, Ser151, Met66 and the aliphatic portion of Arg67. Figure drawn from Protein Data Bank deposition 1SHC. (f) Crystal structure of the IRS-1 PTB domain (green) bound to a phosphorylated segment of the juxtamembrane region of the IR (brown). Figure drawn from Protein Data Bank deposition 5U1M.
