Rational Peptide Design for Inhibition of the KIX–MLL Interaction

May 9, 2023

Title

Rational Peptide Design for Inhibition of the KIX–MLL Interaction

Author

Sato, Nao, Shunji Suetaka, Yuuki Hayashi, and Munehito Arai

Year

2023

Journal

Scientific Reports

Abstract

The kinase-inducible domain interacting (KIX) domain is an integral part of the general transcriptional coactivator CREB-binding protein, and has been associated with leukemia, cancer, and various viral diseases. Hence, the KIX domain has attracted considerable attention in drug discovery and development. Here, we rationally designed a KIX inhibitor using a peptide fragment corresponding to the transactivation domain (TAD) of the transcriptional activator, mixed-lineage leukemia protein (MLL). We performed theoretical saturation mutagenesis using the Rosetta software to search for mutants expected to bind KIX more tightly than the wild-type MLL TAD. Mutant peptides with higher helical propensities were selected for experimental characterization. We found that the T2857W mutant of the MLL TAD peptide had the highest binding affinity for KIX compared to the other 12 peptides designed in this study. Moreover, the peptide had a high inhibitory effect on the KIX–MLL interaction with a half-maximal inhibitory concentration close to the dissociation constant for this interaction. To our knowledge, this peptide has the highest affinity for KIX among all previously reported inhibitors that target the MLL site of KIX. Thus, our approach may be useful for rationally developing helical peptides that inhibit protein–protein interactions implicated in the progression of various diseases.

Instrument

J-805

Keywords

kinase-inducible domain interacting (KIX) domain, inhibitor,peptide drug