TY - JOUR
T1 - Direct interactions among Ret, GDNF and GFRα1 molecules reveal new insights into the assembly of a functional three-protein complex
AU - Amoresano, Angela
AU - Incoronato, Mariarosaria
AU - Monti, Gianluca
AU - Pucci, Piero
AU - De Franciscis, Vittorio
AU - Cerchia, Laura
PY - 2005/6
Y1 - 2005/6
N2 - 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.
AB - 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.
KW - GDNF
KW - MALDI mass spectrometry
KW - Ret
KW - Tyrosine kinase receptor
UR - http://www.scopus.com/inward/record.url?scp=13844252032&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=13844252032&partnerID=8YFLogxK
U2 - 10.1016/j.cellsig.2004.10.012
DO - 10.1016/j.cellsig.2004.10.012
M3 - Article
C2 - 15722196
AN - SCOPUS:13844252032
VL - 17
SP - 717
EP - 727
JO - Cellular Signalling
JF - Cellular Signalling
SN - 0898-6568
IS - 6
ER -