Inhibition of hGrb10 binding to the insulin receptor by functional domain-mediated oligomerization

Lily Q Dong, Sarah Porter, Derong Hu, Feng Liu

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

hGrb10 is a newly identified Src homology 2 (SH2) and pleckstrin homology (PH) domain-containing protein that binds to autophosphorylated receptor tyrosine kinases, including the insulin and insulin-like growth factor receptors. To identify potential downstream proteins that interact with hGrb10, we screened a yeast two-hybrid cDNA library using the full- length hGrb10γ as bait. A fragment of hGrb10, which included the IPS (insert between the PH and SH2 domain) and the SH2 domains, was found to bind with high affinity to the full-length protein. The interaction between the IPS/SH2 domain and the full-length hGrb10 was further confirmed by in vitro glutathione S-transferase fusion protein binding studies. Gel filtration assays showed that hGrb10 underwent tetramerization in mammalian cells. The interaction involved at least two functional domains, the IPS/SH2 region and the PH domain, both of which interacted with the NH2-terminal amino acid sequence of hGrb10γ (hGrb10γΔC, residues 4-414). Competition studies showed that hGrb10γΔC inhibited the binding of hGrb10 to the tyrosine- phosphorylated insulin receptor, suggesting that this region may play a regulatory role in hGrb10/insulin receptor interaction. We present a model for hGrb10 tetramerization and its potential role in receptor tyrosine kinase signal transduction.

Original languageEnglish (US)
Pages (from-to)17720-17725
Number of pages6
JournalJournal of Biological Chemistry
Volume273
Issue number28
DOIs
StatePublished - Jul 10 1998

Fingerprint

Oligomerization
src Homology Domains
Insulin Receptor
Receptor Protein-Tyrosine Kinases
Somatomedin Receptors
Signal transduction
Proteins
Glutathione Transferase
Gene Library
Yeast
Tyrosine
Assays
Fusion reactions
Gels
Cells
Protein Binding
Insulin
Gel Chromatography
Amino Acids
Amino Acid Sequence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibition of hGrb10 binding to the insulin receptor by functional domain-mediated oligomerization. / Dong, Lily Q; Porter, Sarah; Hu, Derong; Liu, Feng.

In: Journal of Biological Chemistry, Vol. 273, No. 28, 10.07.1998, p. 17720-17725.

Research output: Contribution to journalArticle

@article{94db176353e94a5788f0bbc179c481de,
title = "Inhibition of hGrb10 binding to the insulin receptor by functional domain-mediated oligomerization",
abstract = "hGrb10 is a newly identified Src homology 2 (SH2) and pleckstrin homology (PH) domain-containing protein that binds to autophosphorylated receptor tyrosine kinases, including the insulin and insulin-like growth factor receptors. To identify potential downstream proteins that interact with hGrb10, we screened a yeast two-hybrid cDNA library using the full- length hGrb10γ as bait. A fragment of hGrb10, which included the IPS (insert between the PH and SH2 domain) and the SH2 domains, was found to bind with high affinity to the full-length protein. The interaction between the IPS/SH2 domain and the full-length hGrb10 was further confirmed by in vitro glutathione S-transferase fusion protein binding studies. Gel filtration assays showed that hGrb10 underwent tetramerization in mammalian cells. The interaction involved at least two functional domains, the IPS/SH2 region and the PH domain, both of which interacted with the NH2-terminal amino acid sequence of hGrb10γ (hGrb10γΔC, residues 4-414). Competition studies showed that hGrb10γΔC inhibited the binding of hGrb10 to the tyrosine- phosphorylated insulin receptor, suggesting that this region may play a regulatory role in hGrb10/insulin receptor interaction. We present a model for hGrb10 tetramerization and its potential role in receptor tyrosine kinase signal transduction.",
author = "Dong, {Lily Q} and Sarah Porter and Derong Hu and Feng Liu",
year = "1998",
month = "7",
day = "10",
doi = "10.1074/jbc.273.28.17720",
language = "English (US)",
volume = "273",
pages = "17720--17725",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "28",

}

TY - JOUR

T1 - Inhibition of hGrb10 binding to the insulin receptor by functional domain-mediated oligomerization

AU - Dong, Lily Q

AU - Porter, Sarah

AU - Hu, Derong

AU - Liu, Feng

PY - 1998/7/10

Y1 - 1998/7/10

N2 - hGrb10 is a newly identified Src homology 2 (SH2) and pleckstrin homology (PH) domain-containing protein that binds to autophosphorylated receptor tyrosine kinases, including the insulin and insulin-like growth factor receptors. To identify potential downstream proteins that interact with hGrb10, we screened a yeast two-hybrid cDNA library using the full- length hGrb10γ as bait. A fragment of hGrb10, which included the IPS (insert between the PH and SH2 domain) and the SH2 domains, was found to bind with high affinity to the full-length protein. The interaction between the IPS/SH2 domain and the full-length hGrb10 was further confirmed by in vitro glutathione S-transferase fusion protein binding studies. Gel filtration assays showed that hGrb10 underwent tetramerization in mammalian cells. The interaction involved at least two functional domains, the IPS/SH2 region and the PH domain, both of which interacted with the NH2-terminal amino acid sequence of hGrb10γ (hGrb10γΔC, residues 4-414). Competition studies showed that hGrb10γΔC inhibited the binding of hGrb10 to the tyrosine- phosphorylated insulin receptor, suggesting that this region may play a regulatory role in hGrb10/insulin receptor interaction. We present a model for hGrb10 tetramerization and its potential role in receptor tyrosine kinase signal transduction.

AB - hGrb10 is a newly identified Src homology 2 (SH2) and pleckstrin homology (PH) domain-containing protein that binds to autophosphorylated receptor tyrosine kinases, including the insulin and insulin-like growth factor receptors. To identify potential downstream proteins that interact with hGrb10, we screened a yeast two-hybrid cDNA library using the full- length hGrb10γ as bait. A fragment of hGrb10, which included the IPS (insert between the PH and SH2 domain) and the SH2 domains, was found to bind with high affinity to the full-length protein. The interaction between the IPS/SH2 domain and the full-length hGrb10 was further confirmed by in vitro glutathione S-transferase fusion protein binding studies. Gel filtration assays showed that hGrb10 underwent tetramerization in mammalian cells. The interaction involved at least two functional domains, the IPS/SH2 region and the PH domain, both of which interacted with the NH2-terminal amino acid sequence of hGrb10γ (hGrb10γΔC, residues 4-414). Competition studies showed that hGrb10γΔC inhibited the binding of hGrb10 to the tyrosine- phosphorylated insulin receptor, suggesting that this region may play a regulatory role in hGrb10/insulin receptor interaction. We present a model for hGrb10 tetramerization and its potential role in receptor tyrosine kinase signal transduction.

UR - http://www.scopus.com/inward/record.url?scp=0032504161&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032504161&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.28.17720

DO - 10.1074/jbc.273.28.17720

M3 - Article

C2 - 9651371

AN - SCOPUS:0032504161

VL - 273

SP - 17720

EP - 17725

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 28

ER -