Unknown

Dataset Information

0

Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers


ABSTRACT: Chimeric proteins have been widely used to evaluate intracellular protein–protein interactions (PPIs) in living cells with various readouts. By combining an interleukin-3-dependent murine cells and chimeric proteins containing a receptor tyrosine kinase c-kit, we previously established a c-kit-based PPI screening (KIPPIS) system to evaluate and select protein binders. In the KIPPIS components, proteins of interest are connected with a chemically inducible helper module and the intracellular domain of the growth-signaling receptor c-kit, which detects PPIs based on cell proliferation as a readout. In this system, proteins of interest can be incorporated into chimeric proteins without any scaffold proteins, which would be advantageous for evaluating interaction between small peptides/domains. To prove this superiority, we apply KIPPIS to 6 peptide aptamer–polypeptide pairs, which are derived from endogenous, synthetic, and viral proteins. Consequently, all of the 6 peptide aptamer–polypeptide interactions are successfully detected by cell proliferation. The detection sensitivity can be modulated in a helper ligand-dependent manner. The assay results of KIPPIS correlate with the activation levels of Src, which is located downstream of c-kit-mediated signal transduction. Control experiments reveal that KIPPIS clearly discriminates interacting aptamers from non-interacting ones. Thus, KIPPIS proves to be a versatile platform for evaluating the binding properties of peptide aptamers.

SUBMITTER: Kashima D 

PROVIDER: S-EPMC8175380 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC5823606 | biostudies-literature
| S-EPMC4701644 | biostudies-other
| S-EPMC4215913 | biostudies-other
| S-EPMC3519987 | biostudies-literature
| S-EPMC4428161 | biostudies-literature
| S-EPMC6101487 | biostudies-literature
| S-EPMC5859181 | biostudies-literature
| S-EPMC3755880 | biostudies-other
| S-EPMC3722562 | biostudies-literature
2019-09-30 | GSE124865 | GEO