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Negative allosteric modulation of the glucagon receptor by RAMP2.

Kaavya Krishna Kumar1, Evan S O'Brien1, Chris H Habrian1

  • 1Department of Molecular and Cellular Physiology, Stanford University School of Medicine, 279 Campus Drive, Stanford, CA 94305, USA.

Cell
|March 31, 2023
PubMed
Summary

Receptor activity-modifying protein 2 (RAMP2) inhibits glucagon receptor (GCGR) signaling by increasing receptor flexibility. This interaction disrupts GCGR

Keywords:
G-proteinG-protein coupled receptorGPCRHDX-MSallosterycell signalingcryo-EMcryo-electron microscopyglucagon receptorhydrogen-deuterium exchange monitored by mass spectrometryprotein dynamicsreceptor activity-modifying protein RAMPsingle molecule fluorescence resonance energy transfersmFRET

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Receptor activity-modifying proteins (RAMPs) are key regulators of Family B G protein-coupled receptors (GPCRs).
  • The glucagon receptor (GCGR) plays a critical role in maintaining blood sugar homeostasis.

Purpose of the Study:

  • To investigate the interaction between RAMP2 and GCGR.
  • To elucidate the mechanism by which RAMP2 modulates GCGR activity.

Main Methods:

  • Hydrogen-deuterium exchange mass spectrometry (HDX-MS)
  • Somatic cell-surface fluorescence resonance energy transfer (smFRET)
  • Cryo-electron microscopy (cryo-EM)

Main Results:

  • RAMP2 directly interacts with GCGR, broadly inhibiting downstream signaling.
  • RAMP2 enhances local flexibility in the GCGR extracellular domain (ECD) and transmembrane helix 6.
  • Cryo-EM reveals RAMP2-bound GCGR with a disordered ECD and rearranged intracellular activation hallmarks.

Conclusions:

  • RAMP2 acts as a negative allosteric modulator of GCGR.
  • RAMP2 inhibits GCGR by promoting conformational sampling of the extracellular domain, leading to reduced signaling.