From protein-protein complexes to interactomics.

Protein-protein interactions (or PPIs) are key elements for the normal functioning of a living cell. A large description of the protein interactomics field is given in this review where different aspects will be discussed. We first give an introduction of the different large scale experimental approaches from yeast two-hybrid to mass spectrometry used to discover PPIs and build protein interaction maps. Single PPI validation techniques such as co-immunoprecipitation or fluorescence methods are then presented as they are more and more integrated in global PPI discovery strategy. Data from different experimental sets are compared and an assessment of the different large scale technologies is presented. Bioinformatics tools can also predict with a good accuracy PPIs in silico, PPIs databases are now numerous and topological analysis has led to interesting insights into the nature of network connection. Finally, PPI, as an association of two proteins, has been structurally characterized for many protein complexes and is largely discussed throughout existing examples. The results obtained so far already provide the biologist with a large set of structured data from which knowledge on pathways and associated protein function can be extracted.

Small-molecule inhibitor of USP7/HAUSP ubiquitin protease stabilizes and activates p53 in cells.

Deregulation of the ubiquitin/proteasome system has been implicated in the pathogenesis of many human diseases, including cancer. Ubiquitin-specific proteases (USP) are cysteine proteases involved in the deubiquitination of protein substrates. Functional connections between USP7 and essential viral proteins and oncogenic pathways, such as the p53/Mdm2 and phosphatidylinositol 3-kinase/protein kinase B networks, strongly suggest that the targeting of USP7 with small-molecule inhibitors may be useful for the treatment of cancers and viral diseases. Using high-throughput screening, we have discovered HBX 41,108, a small-molecule compound that inhibits USP7 deubiquitinating activity with an IC(50) in the submicromolar range. Kinetics data indicate an uncompetitive reversible inhibition mechanism. HBX 41,108 was shown to affect USP7-mediated p53 deubiquitination in vitro and in cells. As RNA interference-mediated USP7 silencing in cancer cells, HBX 41,108 treatment stabilized p53, activated the transcription of a p53 target gene without inducing genotoxic stress, and inhibited cancer cell growth. Finally, HBX 41,108 induced p53-dependent apoptosis as shown in p53 wild-type and null isogenic cancer cell lines. We thus report the identification of the first lead-like inhibitor against USP7, providing a structural basis for the development of new anticancer drugs.