RT Journal
A1 Colland, Frédéric
A1 Jacq, Xavier
A1 Trouplin, Virginie
A1 Mougin, Christelle
A1 Groizeleau, Caroline
A1 Hamburger, Alexandre
A1 Meil, Alain
A1 Wojcik, Jérôme
A1 Legrain, Pierre
A1 Gauthier, Jean-Michel
T1 Functional Proteomics Mapping of a Human Signaling Pathway
JF Genome Research 
JO Genome Research 
YR 2004 
FD July 01 
VO 14 
IS 7 
SP 1324 
OP 1332 
DO 10.1101/gr.2334104 
UL http://genome.cshlp.org/content/14/7/1324.abstract 
AB Access to the human genome facilitates extensive functional proteomics studies. Here, we present an integrated approach combining large-scale protein interaction mapping, exploration of the interaction network, and cellular functional assays performed on newly identified proteins involved in a human signaling pathway. As a proof of principle, we studied the Smad signaling system, which is regulated by members of the transforming growth factor β (TGFβ) superfamily. We used two-hybrid screening to map Smad signaling protein–protein interactions and to establish a network of 755 interactions, involving 591 proteins, 179 of which were poorly or not annotated. The exploration of such complex interaction databases is improved by the use of PIMRider, a dedicated navigation tool accessible through the Web. The biological meaning of this network is illustrated by the presence of 18 known Smad-associated proteins. Functional assays performed in mammalian cells including siRNA knock-down experiments identified eight novel proteins involved in Smad signaling, thus validating this integrated functional proteomics approach.