Reference - Detail
| RRC ID | 45241 |
|---|---|
| Author | Mabe S, Nagamune T, Kawahara M. |
| Title | Detecting protein-protein interactions based on kinase-mediated growth induction of mammalian cells. |
| Journal | Sci Rep |
| 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 of mammalian cells. In this method, target proteins are fused to the intracellular domain of c-kit (c-kit ICD) and expressed in interleukin-3-dependent mammalian cells. The PPI induces dimerization and activation of c-kit ICDs, which leads to cell growth in the absence of interleukin-3. Using this system, we successfully detected the ligand-dependent homo-interaction of FKBPF36V and hetero-interaction of FKBP and FRBT2098L, as well as the constitutive interaction between MDM2 and a known peptide inhibitor. Intriguingly, cells expressing high-affinity peptide chimeras are selected from the mixture of the cell populations dominantly expressing low-affinity peptide chimeras. These results indicate that this method can detect PPIs with low background levels and is suitable for peptide inhibitor screening. |
| Volume | 4 |
| Pages | 6127 |
| Published | 2014-8-19 |
| DOI | 10.1038/srep06127 |
| PII | srep06127 |
| PMID | 25135216 |
| PMC | PMC4137342 |
| MeSH | Amino Acid Substitution Animals Cell Line Cell Proliferation / drug effects Dimerization Mice Peptides / chemistry Peptides / metabolism Protein Interaction Domains and Motifs Protein Interaction Mapping / methods* Proto-Oncogene Proteins c-kit / chemistry Proto-Oncogene Proteins c-kit / genetics Proto-Oncogene Proteins c-kit / metabolism* Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors Proto-Oncogene Proteins c-mdm2 / metabolism Tacrolimus / analogs & derivatives Tacrolimus / chemistry Tacrolimus / metabolism Tacrolimus / pharmacology Tacrolimus Binding Proteins / chemistry Tacrolimus Binding Proteins / genetics Tacrolimus Binding Proteins / metabolism Tumor Suppressor Protein p53 / chemistry Tumor Suppressor Protein p53 / metabolism |
| IF | 3.998 |
| Times Cited | 9 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Resource | |
| Human and Animal Cells | Ba/F3(RCB0805) |