A membrane protein / signaling protein interaction network for Arabidopsis version AMPv2

• DOI: 10.3389/fphys.2010.00024
• Publication Date: 2010-09-22T13:37:42Z
• Authors: Sylvie Lalonde
• Keywords: Interactome

Interactions between membrane proteins and the soluble fraction are essential for signal transduction regulating nutrient transport. To gain insights into membrane-based interactome, 3,852 open reading frames (ORFs) out of a target list 8,383 representing signaling from Arabidopsis thaliana were cloned Gateway compatible vector. The mating-based split-ubiquitin system was used to screen potential protein-protein interactions (pPPIs) among 490 ORFs. A binary robotic 142 receptor-like kinases, 72 transporters, 57 protein kinases phosphatases, 40 glycosyltransferases, 95 various functions 89 with unknown function detected 387 90,370 possible PPIs. secondary confirmed 343 (of 387) pPPIs 179 proteins, yielding scale-free network (r2=0.863). Eighty transmembrane (RLK) tested positive, identifying three homomers, 63 heteromers 80 other proteins. Thirty-one RLK interactors (including RLKs) had previously been found be phosphorylated; thus may substrates respective RLKs. None described here reported in major interactome databases, including G protein-coupled receptors, phospholipase C, AMT ammonium transporters. Two RLKs as putative AMT1;1 independently using split luciferase assay protoplasts. These involved ammonium-dependent phosphorylation C-terminus regulation uptake activity. screening method established will enable systematic analysis fungi, plants metazoa.