Single-molecule FRET and molecular dynamics simulations reveal early activation steps of MET receptor by Listeria monocytogenes

• Published in: bioRxiv
• Main Authors: Li, Yunqing, Arghittu, Serena M, Dietz, Marina S, Hasse, Daniel, Ferraris, Davide M, Freund, Petra, Barth, Hans-Dieter, Niemann, Harmut H, Covino, Roberto, Heilemann, Mike
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 23.12.2023
• Subjects: c-Met protein; Cell migration; Cell proliferation; Cell surface receptors; Cell survival; Internalin; Internalization; Kinases; Listeria; Listeria monocytogenes; Pathogens; Protein-tyrosine kinase receptors; Simulation
• DOI: 10.1101/2023.12.22.572978

The human growth factor receptor MET is a receptor tyrosine kinase involved in cell proliferation, migration, and survival. MET is also hijacked by the intracellular pathogen Listeria monocytogenes. Its invasion protein, internalin B (InlB), binds to MET and promotes the formation of a signaling dimer that triggers the internalization of the pathogen. Here, we use a combination of structural biology, modeling, molecular dynamics simulations, and in situ single-molecule Foerster resonance energy transfer (smFRET) experiments to elucidate the early events in MET activation by Listeria. Simulations show that InlB binding stabilizes MET in a conformation that promotes dimer formation. smFRET identifies the organization of the in situ signaling dimer. Further MD simulations of the dimer model are in quantitative agreement with smFRET. We accurately describe the structural dynamics underpinning an important cellular event and introduce a powerful methodological pipeline applicable to studying the activation of other plasma membrane receptors.Competing Interest StatementThe authors have declared no competing interest.