The N terminus of adhesion G protein-coupled receptor GPR126/ADGRG6 as allosteric force integrator

Mitgau, Jakob; Franke, Julius; Schinner, Camilla; Stephan, Gabriele; Berndt, Sandra; Placantonakis, Dimitris G.; Spindler, Volker; Wilde, Caroline; Liebscher, Ines

doi:10.1101/2021.09.13.460127

Cited by 3 publications

(2 citation statements)

References 61 publications

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“…While these studies did not specifically test the effects of force-mediated dissociation on aGPCR signaling, the dissociation times for the Lphn3 GAIN domain measured in this study are consistent with the timescales reported in the literature for signaling via latrophilins and other aGPCRs. Previous studies on latrophilins and other aGPCRs have established a link between force and signaling 25,26 , but the work presented here represents to the first study to our knowledge that characterizes forces involved in aGPCR dissociation at the molecular level. In the context of synaptogenesis, it remains to be tested whether bonds between latrophilins and ligands such as teneurins or FLRTs can support the forces necessary to dissociate latrophilins at the timescales that we observe.…”

Section: Discussionmentioning

confidence: 93%

Piconewton forces mediate GAIN domain dissociation of the latrophilin-3 adhesion GPCR

Zhong

Lee

Vachharajani

et al. 2023

Preprint

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Latrophilins are adhesion G-protein coupled receptors (aGPCRs) that control excitatory synapse formation. aGPCRs, including latrophilins, are autoproteolytically cleaved at their GPCR-Autoproteolysis Inducing (GAIN) domain, but the two resulting fragments remain associated on the cell surface. It is thought that force-mediated dissociation of the fragments exposes a peptide that activates G-protein signaling of aGPCRs, but whether GAIN domain dissociation can occur on biologically relevant timescales and at physiological forces is unknown. Here, we show using magnetic tweezers that physiological forces dramatically accelerate the dissociation of the latrophilin-3 GAIN domain. Forces in the 1-10 pN range were sufficient to dissociate the GAIN domain on a seconds-to-minutes timescale, and the GAIN domain fragments reversibly reassociated after dissociation. Thus, mechanical force may be a key driver of latrophilin signaling during synapse formation, suggesting a physiological mechanism by which aGPCRs may mediate mechanically-induced signal transduction.

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Section: Discussionmentioning

confidence: 93%

Piconewton forces mediate GAIN domain dissociation of the latrophilin-3 adhesion GPCR

Zhong

Lee

Vachharajani

et al. 2023

Preprint

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“…Third, our studies evaluate the interaction of the tethered agonist with ECL2 in a context devoid of the NTF. In a setting of peptide- or antibody-based activation without NTF removal, exemplified recently by Mitgau et al, 2022 for GPR126, these tethered agonist/ECL interactions may be different.…”

Section: Discussionmentioning

confidence: 99%

Regulation of pulmonary surfactant by the adhesion GPCR GPR116/ADGRF5 requires a tethered agonist-mediated activation mechanism

Bridges

Safina

Pirard

et al. 2022

eLife

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The mechanistic details of the tethered agonist mode of activation for the adhesion GPCR ADGRF5/GPR116 has not been completely deciphered. We set out to investigate the physiologic importance of autocatalytic cleavage upstream of the agonistic peptide sequence, an event necessary for NTF displacement and subsequent receptor activation. To examine this hypothesis, we characterized tethered agonist-mediated activation of GPR116 in vitro and in vivo. A knock-in mouse expressing a non-cleavable GPR116 mutant phenocopies the pulmonary phenotype of GPR116 knock-out mice, demonstrating that tethered agonist-mediated receptor activation is indispensable for function in vivo. Using site-directed mutagenesis and species swapping approaches we identified key conserved amino acids for GPR116 activation in the tethered agonist sequence and in extracellular loops 2/3 (ECL2/3). We further highlight residues in transmembrane7 (TM7) that mediate stronger signaling in mouse versus human GPR116 and recapitulate these findings in a model supporting tethered agonist:ECL2 interactions for GPR116 activation.

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Step-wise mechanical unfolding and dissociation of the GAIN domains of ADGRG1/GPR56, ADGRL1/Latrophilin-1 and ADGRB3/BAI3: insights into the mechanical activation hypothesis of adhesion G protein-coupled receptors

Huang

Tang

et al. 2023

Preprint

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Adhesion G protein-coupled receptors (aGPCRs) are a large family within the superfamily of G protein-coupled receptors involved in various physiological processes. One unique feature of aGPCRs is their long N-terminal extracellular regions (ECRs), which contain adhesive domains and a GPCR autoproteolysis-inducing (GAIN) domain. This GAIN domain promotes autoproteolytic cleavage of aGPCRs into N- and C-terminal fragments (NTF, CTF, respectively) after receptor biosynthesis. aGPCR signaling involves an interplay between the NTF and CTF that can be mechanically activated or modulated. However, how force affects the conformation/structure of the GAIN domain as a central structural element in aGPCR activation remains largely unknown. In this study, we investigated the mechanical stability of the GAIN domains of three aGPCRs from subfamilies B, G and L at a loading rate of 1 pN/s. Our findings demonstrate that the GAIN domains can be destabilized by forces from a few to 20 piconewtons (pN). Specifically, for the autocleaved aGPCRs, ADGRG1/GPR56 and ADGRL1/Latrophilin-1, forces over this range can cause detachment of the GAIN domain from the membrane-proximal Stachel element, which serves as an endogenous tethered agonist to aGPCRs, typically preceded with GAIN domain unfolding. For the non-cleavable aGPCR ADGRB3/BAI3, the GAIN domain undergoes complex mechanical unfolding over a similar force range. We also demonstrate that detachment of the GAIN domain can take place during cell migration, provided that the linkage between aGPCR and extracellular matrix is sufficiently stable. These results suggest that both structural stability of the GAIN domain and NTF/CTF dissociation are sensitive to physiological ranges of tensile forces, providing insights into the mechanical activation hypothesis of aGPCRs.

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The N terminus of adhesion G protein-coupled receptor GPR126/ADGRG6 as allosteric force integrator

Cited by 3 publications

References 61 publications

Piconewton forces mediate GAIN domain dissociation of the latrophilin-3 adhesion GPCR

Piconewton forces mediate GAIN domain dissociation of the latrophilin-3 adhesion GPCR

Regulation of pulmonary surfactant by the adhesion GPCR GPR116/ADGRF5 requires a tethered agonist-mediated activation mechanism

Step-wise mechanical unfolding and dissociation of the GAIN domains of ADGRG1/GPR56, ADGRL1/Latrophilin-1 and ADGRB3/BAI3: insights into the mechanical activation hypothesis of adhesion G protein-coupled receptors

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