RAMP3 determines rapid recycling of atypical chemokine receptor-3 for guided angiogenesis

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 116; no. 48; pp. 24093 - 24099
• Main Authors: Mackie, Duncan I., Nielsen, Natalie R., Harris, Matthew, Singh, Smriti, Davis, Reema B., Dy, Danica, Ladds, Graham, Caron, Kathleen M.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 26.11.2019
• Subjects: Adrenomedullin; Allosteric properties; Angiogenesis; Biological Sciences; Bioluminescence; Bioluminescence Resonance Energy Transfer Techniques; Cell adhesion & migration; Cell Movement; Cell surface; Chaperones; Chemokine receptors; Chemokines; Clonal deletion; Coevolution; Coupling (molecular); CXCL12 protein; Energy transfer; G protein-coupled receptors; HEK293 Cells; Humans; Leukocyte migration; Lysosomes - metabolism; Membrane vesicles; Modulators; Molecular interactions; Neovascularization, Physiologic; Protein transport; Proteins; Ramps; Receptor activity modifying proteins; Receptor Activity-Modifying Protein 3 - genetics; Receptor Activity-Modifying Protein 3 - metabolism; Receptor Activity-Modifying Protein 3 - physiology; Receptor mechanisms; Receptors; Receptors, CCR3 - metabolism; Receptors, CXCR - metabolism; Retina; SDF-1 protein; Signal Transduction; Signaling; chemokine receptors; endosomal sorting; guided cell migration; receptor-activity–modifying proteins; G-protein–coupled receptors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1905561116

Receptor-activity–modifying proteins (RAMPs) are single transmembrane-spanning proteins which serve as molecular chaperones and allosteric modulators of G-protein–coupled receptors (GPCRs) and their signaling pathways. Although RAMPs have been previously studied in the context of their effects on Family B GPCRs, the coevolution of RAMPs with many GPCR families suggests an expanded repertoire of potential interactions. Using bioluminescence resonance energy transfer-based and cell-surface expression approaches, we comprehensively screen for RAMP interactions within the chemokine receptor family and identify robust interactions between RAMPs and nearly all chemokine receptors. Most notably, we identify robust RAMP interaction with atypical chemokine receptors (ACKRs), which function to establish chemotactic gradients for directed cell migration. Specifically, RAMP3 association with atypical chemokine receptor 3 (ACKR3) diminishes adrenomedullin (AM) ligand availability without changing G-protein coupling. Instead, RAMP3 is required for the rapid recycling of ACKR3 to the plasma membrane through Rab4-positive vesicles following either AM or SDF-1/CXCL12 binding, thereby enabling formation of dynamic spatiotemporal chemotactic gradients. Consequently, genetic deletion of either ACKR3 or RAMP3 in mice abolishes directed cell migration of retinal angiogenesis. Thus, RAMP association with chemokine receptor family members represents a molecular interaction to control receptor signaling and trafficking properties.