Activation Pathways of Neurotensin Receptor 1 Elucidated Using Statistical Machine Learning

: Neurotensin receptor 1 (NTSR1) is a G-protein coupled receptor (GPCR) that mediates many biological processes through its interaction with the neurotensin (NTS) peptide. The NTSR1 protein is a clinically significant target as it is involved in the proliferation of cancer cells. Understanding the activation mechanism of NTSR1 is an important prerequisite for exploring the therapeutic potential of targeting NTSR1 and the development of drug molecules specific to NTSR1. Previous studies have been aimed at elucidating the structure of NTSR1 in the active and inactive conformations; however, the intermediate molecular pathway for NTSR1 activation dynamics is largely unknown. In this study, we performed extensive molecular dynamics (MD) simulations of the NTSR1 protein and analyzed its kinetic conformational changes to determine the microswitches that drive NTSR1 activation. To biophysically interpret the high-dimensional simulation trajectories, we used Markov state models and machine learning to elucidate the important and detailed conformational changes in NTSR1. Through the analysis of identified microswitches, we propose a mechanistic pathway for NTSR1 activation.