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© Michel Huerre
Coupe histologique de foie de patient atteint d'une hépatite C chronique active avec infiltrat (opacité) folliculaire (lymphocytes) (Grossissement X 400). L'hépatite C chronique peut être à l'origine de cirrhose et cancer du foie
Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nature methods - 29 Nov 2011

Cassonnet P, Rolloy C, Neveu G, Vidalain PO, Chantier T, Pellet J, Jones L, Muller M, Demeret C, Gaud G, Vuillier F, Lotteau V, Tangy F, Favre M, Jacob Y

Link to Pubmed [PMID] – 22127214

Nat. Methods 2011 Dec;8(12):990-2

Capture d’écran 2015-09-03 à 18.19.08

Mapping protein-protein interactions (PPIs) has proven instrumental in functional proteomics. Large PPI databases have been established by literature mining or using high-throughput screening methods; this information requires cross-validation to increase coverage and accuracy. Previously published papers have argued for the use of defined reference sets for the benchmarking of screening methods for PPIs. Here we describe a high-throughput system for orthogonal validation of PPI datasets, and we benchmark its performance against newly curated and previously reported reference sets. Based on a previously reported PPI-detection assay relying on complementation of split Gaussia princeps luciferase, we engineered two mammalian expression vectors for high-throughput PPI detection. In this assay, we monitored PPIs by measuring interaction-mediated normalized luminescence ratio (NLR). We benchmarked the accuracy and sensitivity of this system using an a priori ‘negative’ or non-interacting set of 100 random human protein pairs (the random reference set, RRS) and a positive or interacting set composed of 143 PPIs from NF-kB, IFN-a/b and TGF-b pathways supported in the literature by at least three different experimental methods4 (the positive reference set, PRS).

http://www.ncbi.nlm.nih.gov/pubmed/22127214