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bioRxiv · 10.1101/2025.08.04.668378

Mechanism of Tethered Agonist Binding to an Adhesion G-Protein-Coupled Receptor

Joshi, K.·Miao, Y.
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Abstract

Adhesion GPCRs (ADGRs) contain a GPCR autoproteolysis-inducing (GAIN) domain that is proximal to the receptor N-terminus and undergoes autoproteolysis at a highly conserved GPCR proteolysis (GPS) site to generate the N-terminal fragment (NTF) and transmembrane C-terminal fragment (CTF). Dissociation of NTF reveals a peptide tethered agonist (TA) which is responsible for the activation of the ADGRs. The NTF-bound ADGRs contain the encrypted TA that assumes a {beta}-strand configuration within the GAIN domain, which is markedly different from a U-shaped -helical configuration of TA in the active cryo-EM structures of ADGRs. However, how the TA dramatically changes its configuration and binds to the ADGR CTF remains unknown. In this study, we have performed all-atom enhanced sampling simulations using a novel Peptide-Gaussian accelerated Molecular Dynamics (Pep-GaMD) method on TA binding to the ADGRD1. The Pep-GaMD simulations captured spontaneous binding of the TA into orthosteric pocket of the ADGRD1 and its large conformational transition from the extended {beta} strand to the U-shaped helical configuration. We were able to identify important low-energy conformations of the TA in the binding pathway, as well as different active and inactive states of ADGRD1, in the presence and absence of the Gs protein. Therefore, our Pep-GaMD simulations have revealed dynamic mechanism of the TA binding to an ADGR, which will facilitate rational design of peptide regulators of ADGRs. TOC Graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/668378v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@15e1fborg.highwire.dtl.DTLVardef@a73738org.highwire.dtl.DTLVardef@1a63d2org.highwire.dtl.DTLVardef@8f848f_HPS_FORMAT_FIGEXP M_FIG C_FIG Peptide-Gaussian accelerated Molecular Dynamics (Pep-GaMD) simulations remarkably captured spontaneous binding of tethered agonist (TA) into the orthosteric pocket of adhesion G-protein-coupled receptor (ADGRD1) and large conformational transition of the TA from an extended {beta}-strand to a U-shaped -helical configuration. We were able to identify important low-energy conformations of the TA in the binding pathway and different states of the receptor from the Pep-GaMD simulations.

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Joshi, K., Miao, Y.. 2025-08-04. Mechanism of Tethered Agonist Binding to an Adhesion G-Protein-Coupled Receptor. https://doi.org/10.1101/2025.08.04.668378

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