Direct interactions among Ret, GDNF and GFRα1 molecules reveal new insights into the assembly of a functional three-protein complex

Angela Amoresano, Mariarosaria Incoronato, Gianluca Monti, Piero Pucci, Vittorio De Franciscis, Laura Cerchia

Research output: Contribution to journalArticlepeer-review

Abstract

The glial-cell-line-derived neurotrophic factor (GDNF) ligand activates the Ret receptor through the assembly of a multiprotein complex, including the GDNF family receptor α1 (GFRα1) molecule. Given the neuroprotective role of GDNF, there is an obvious need to precisely identify the structural regions engaged in direct interactions between the three molecules. Here, we combined a functional approach for Ret activity (in PC12 cells) to cross-linking experiments followed by MS-MALDI to study the interactions among the purified extracellular region of the human Ret, GDNF and GFRα1 molecules. This procedure allowed us to identify distinct regions of Ret that are physically engaged in the interaction with GDNF and GFRα1. The lack of these regions in a recombinant Ret form results in the failure of both structural and functional binding of Ret to GFRα1/GDNF complex. Furthermore, a model for the assembly of a transducing-competent Ret complex is suggested.

Original languageEnglish
Pages (from-to)717-727
Number of pages11
JournalCellular Signalling
Volume17
Issue number6
DOIs
Publication statusPublished - Jun 2005

Keywords

  • GDNF
  • MALDI mass spectrometry
  • Ret
  • Tyrosine kinase receptor

ASJC Scopus subject areas

  • Cell Biology

Fingerprint Dive into the research topics of 'Direct interactions among Ret, GDNF and GFRα1 molecules reveal new insights into the assembly of a functional three-protein complex'. Together they form a unique fingerprint.

Cite this