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Imaging Denatured Collagen Strands In vivo and Ex vivo via Photo-triggered Hybridization of Caged Collagen Mimetic Peptides
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Pinpointing phosphotyrosine-dependent interactions downstream of the collagen receptor DDR1

Diana H H Koo1, Catherine McFadden, Yun Huang

  • 1Department of Laboratory Medicine and Pathobiology, University of Toronto, Medical Sciences Building, Room 7334, 1 King's College Circle, Toronto, Ont., Canada M5S 1A8.

FEBS Letters
|December 13, 2005
PubMed

Insights

Collagen-activated DDR1 (discoidin domain receptor 1) interacts with Nck2 and Shp-2 proteins. Ablation of DDR1 disrupts Nck2 localization, impacting mammary gland alveologenesis.

Area of Science:

  • Cell biology
  • Molecular signaling
  • Biochemistry

Background:

  • Receptor tyrosine kinases (RTKs) like DDR1 play crucial roles in cellular processes.
  • DDR1 activation by collagen leads to sustained phosphorylation, but downstream signaling pathways remain largely uncharacterized.
  • Understanding DDR1's interactors is vital for deciphering its biological functions.

Purpose of the Study:

  • To identify and characterize the downstream mediators of collagen-induced DDR1 activation.
  • To elucidate the specific binding sites and interactions between DDR1 and its associated proteins.
  • To investigate the functional consequences of DDR1-mediated signaling in mammary gland development.

Main Methods:

  • Phosphopeptide mapping to identify tyrosine phosphorylation sites on DDR1.
  • Site-directed mutagenesis to confirm phosphorylation sites and binding interactions.
  • Co-immunoprecipitation assays to detect protein-protein interactions.
  • Analysis of DDR1-deficient mouse mammary gland tissue.

Main Results:

  • Multiple novel tyrosine phosphorylation sites were identified within DDR1.
  • Nck2 and Shp-2 were identified as key SH2 domain-containing proteins binding to DDR1 in a collagen-dependent manner.
  • The Shp-2 binding site was mapped to tyrosine-740 within an ITIM-consensus sequence on DDR1.
  • Ablation of DDR1 in mouse mammary glands led to delocalized Nck2 expression.

Conclusions:

  • DDR1 directly interacts with Nck2 and Shp-2 following collagen activation.
  • The DDR1-Shp-2 interaction involves a specific ITIM motif.
  • Disruption of the DDR1-Nck2 interaction contributes to defects in mammary gland alveologenesis.

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