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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
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Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

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The interactome of a PTB domain-containing adapter protein, Odin, revealed by SILAC

Jun Zhong1, Raghothama Chaerkady, Kumaran Kandasamy

  • 1McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA.

Journal of Proteomics
|November 18, 2010
PubMed

Insights

Odin, a negative regulator of growth factor signaling, interacts with 14-3-3 proteins and other novel partners. This study identifies Odin

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Signal Transduction

Background:

  • Signal transduction pathways require precise regulation by positive and negative regulators.
  • Odin (ANKRD1A) was previously identified as a negative regulator of growth factor signaling.
  • The precise mechanisms of Odin's regulatory function were not fully understood.

Purpose of the Study:

  • To elucidate the mechanisms by which Odin negatively regulates growth factor signaling.
  • To systematically identify Odin-interacting proteins using a proteomic approach.

Main Methods:

  • Stable Isotope Labeling by Amino acids in Cell culture (SILAC) strategy was employed for proteomic analysis.
  • Co-immunoprecipitation experiments were used to confirm identified interactions.
  • Literature-based network analysis was performed on Odin interacting partners.

Main Results:

  • Eighteen molecules were identified as specifically associated with Odin in a protein complex.
  • The complete family of 14-3-3 proteins was found to complex with Odin.
  • Novel Odin interactors include CD2-associated protein, SH3 domain kinase binding protein 1, and DAB2 interacting protein.
  • Eight of the eighteen identified interactions were confirmed via co-immunoprecipitation.
  • Network analysis indicated Odin partners are involved in diverse cellular processes, including receptor endocytosis.

Conclusions:

  • Odin interacts with a comprehensive set of 14-3-3 proteins and novel partners.
  • These interactions provide mechanistic insights into Odin's role as a negative regulator of growth factor signaling.
  • Odin's interacting partners are implicated in key cellular processes such as receptor endocytosis.

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