Detecting protein–protein interactions based on kinase-mediated growth induction of mammalian cells

0301 basic medicine 0303 health sciences Proto-Oncogene Proteins c-mdm2 Article Tacrolimus 6. Clean water Cell Line 3. Good health Tacrolimus Binding Proteins Mice Proto-Oncogene Proteins c-kit 03 medical and health sciences Amino Acid Substitution Protein Interaction Mapping Animals Protein Interaction Domains and Motifs Tumor Suppressor Protein p53 Peptides Dimerization Cell Proliferation
DOI: 10.1038/srep06127 Publication Date: 2014-08-19T09:04:48Z
ABSTRACT
Detection of protein–protein interactions (PPIs) is important for understanding numerous processes in mammalian cells; however, existing PPI detection methods often give significant background signals. Here, we propose a novel PPI-detection method based on kinase-mediated growth induction cells. In this method, target proteins are fused to the intracellular domain c-kit (c-kit ICD) and expressed interleukin-3-dependent The induces dimerization activation ICDs, which leads cell absence interleukin-3. Using system, successfully detected ligand-dependent homo-interaction FKBPF36V hetero-interaction FKBP FRBT2098L, as well constitutive interaction between MDM2 known peptide inhibitor. Intriguingly, cells expressing high-affinity chimeras selected from mixture populations dominantly low-affinity chimeras. These results indicate that can detect PPIs with low levels suitable inhibitor screening.
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