Development of an RHEB-Targeting Peptide To Inhibit mTORC1 Kinase Activity

Raef Shams; Yoshihiro Ito; Hideyuki Miyatake

doi:10.1021/acsomega.2c01865

Development of an RHEB-Targeting Peptide To Inhibit mTORC1 Kinase Activity

ACS Omega. 2022 Jun 24;7(27):23479-23486. doi: 10.1021/acsomega.2c01865. eCollection 2022 Jul 12.

Authors

Raef Shams^{1

2}, Yoshihiro Ito^{1

3}, Hideyuki Miyatake^{2

3}

Affiliations

¹ Emergent Bioengineering Materials Research Team, RIKEN Center for Emergent Matter Science, Wako, Saitama 351-0198, Japan.
² Department of Life Science, Graduate School of Science and Engineering, Saitama University, Saitama City, Saitama 338-8570, Japan.
³ Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan.

Abstract

In cancer, the mechanistic/mammalian target of rapamycin complex-1 (mTORC1) is hyperactivated to promote survival under adverse conditions. The kinase activity of mTORC1 is activated by small-GTPase RHEB-GTP. Therefore, a new modality to inhibit mTORC1 activity has emerged, through intercepting RHEB. However, due to the relatively large contact area involved in the interaction between RHEB and mTORC1, facilitating this inhibition through small molecules has been challenging. Here, we report the development of a peptide that can inhibit the RHEB-mTORC1 interaction. The peptide, P1_WT, was designed based on the α-helix (aa 101-115) of the N-heat domain of mTOR to interact with switch II of RHEB. P1_WT bound to RHEB (K _D = 0.14 μM) and inhibited RHEB-mTOR^N-heat interaction (IC₅₀ = 0.33 μM) in vitro. Consequently, P1_WT inhibited mTORC1 activity at a sub-micromolar level (IC₅₀ ∼ 0.3 μM). P1_WT was predicted to be cell-permeable due to the rich content of arginine (23%), enhancing the intracellular translocation. These results show that P1_WT is a potential compound to further develop inhibitors for mTORC1 by intercepting RHEB from mTORC1.