PMID- 10340378
OWN - NLM
STAT- MEDLINE
DCOM- 19990608
LR  - 20091119
IS  - 0950-9232 (Print)
IS  - 0950-9232 (Linking)
VI  - 18
IP  - 20
DP  - 1999 May 20
TI  - Re-engineering the target specificity of the insulin receptor by modification of 
      a PTB domain binding site.
PG  - 3071-5
AB  - Shc and IRS-1 (and their relatives) are cytoplasmic docking proteins that possess
      phosphotyrosine-binding (PTB) domains, through which they bind specific activated
      receptor tyrosine kinases (RTK). The subsequent phosphorylation of Shc or IRS-1
      creates binding sites for the SH2 domains of multiple signaling proteins, leading
      to the activation of intracellular biochemical pathways. The PTB domains of Shc
      and IRS-1 both recognize autophosphorylation sites in RTKs with the consensus
      sequence NPXpY, but show distinct abilities to bind stably to RTKs such as the
      TrkA nerve growth factor receptor and the insulin receptor. In vitro analysis has
      suggested that residues N-terminal to the NPXpY motif may determine the affinity 
      with which phosphopeptide ligands are recognized by the Shc and IRS-1 PTB
      domains. Unlike IRS-1, the Shc PTB domain binds poorly to the insulin-receptor
      (IR) beta subunit in vitro, owing to its low affinity for the NPXpY
      autophosphorylation site at Tyr 960 of the IR. As a consequence, Shc does not
      bind stably to the activated IR in cells. We show that substitution of Ser 955,
      five residues N-terminal to the Tyr 960 autophosphorylation site (the -5
      position), with Ile alters the target specificity of the IR such that it stably
      associates with Shc in insulin-stimulated cells. A triple substitution of the -5,
      -8 and -9 residues relative to Tyr 960 of the IR to the corresponding amino acids
      found in the Shc PTB domain binding site of TrkA results in even stronger binding
      of the IR to Shc in vivo. The variant IRs with enhanced ability to bind Shc
      showed an increased ability to activate the MAPK pathway in response to insulin
      stimulation. These results demonstrate that subtle differences in residues
      N-terminal to NPXpY autophosphorylation sites determine the ability of RTKs to
      bind specific PTB domain proteins in vivo, and thus modify the signaling
      properties of activated receptors.
FAU - van der Geer, P
AU  - van der Geer P
AD  - Department of Chemistry and Biochemistry, University of California, San Diego, La
      Jolla 92093-0601, USA.
FAU - Wiley, S
AU  - Wiley S
FAU - Pawson, T
AU  - Pawson T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Oncogene
JT  - Oncogene
JID - 8711562
RN  - 0 (Insulin Receptor Substrate Proteins)
RN  - 0 (Phosphoproteins)
RN  - 21820-51-9 (Phosphotyrosine)
RN  - EC 2.7.10.1 (Receptor, Insulin)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - Blotting, Western
MH  - COS Cells
MH  - Calcium-Calmodulin-Dependent Protein Kinases/metabolism
MH  - Insulin Receptor Substrate Proteins
MH  - Phosphoproteins/metabolism
MH  - Phosphorylation
MH  - Phosphotyrosine/*metabolism
MH  - Precipitin Tests
MH  - Receptor, Insulin/chemistry/*metabolism
MH  - Signal Transduction
MH  - src Homology Domains
EDAT- 1999/05/26 00:00
MHDA- 1999/05/26 00:01
CRDT- 1999/05/26 00:00
PHST- 1999/05/26 00:00 [pubmed]
PHST- 1999/05/26 00:01 [medline]
PHST- 1999/05/26 00:00 [entrez]
AID - 10.1038/sj.onc.1202879 [doi]
PST - ppublish
SO  - Oncogene. 1999 May 20;18(20):3071-5. doi: 10.1038/sj.onc.1202879.