Microglia-mediated neuroinflammation and α-synuclein (α-syn) aggregation, both as pathological hallmarks of Parkinson’s disease (PD), crosstalk to exacerbate degeneration of dopaminergic neurons and PD progression. However, the mechanism underlying their interaction is poorly understood, which obstructs effective therapeutic inhibition of α-syn-induced neuroinflammation. Here, we initiate from structure-based interaction predictions and find that receptor for advanced glycation end products (RAGE) serves as a receptor of α-syn fibrils on microglia. Results of nuclear magnetic resonance (NMR) spectroscopy and mutagenesis validate that the V domain of RAGE that contains an alkaline surface can bind with acidic C-terminal residues of α-syn. Furthermore, the binding of α-syn fibrils with RAGE induces neuroinflammation, which is blocked by both genetic depletion of RAGE and inhibitor FPS-ZM1. Our work shows the important role, as well as the structural mechanism, of RAGE in mediating the inflammatory response of microglia to α-syn fibrils, which may help to establish effective therapeutic strategies to alleviate α-syn-induced neuroinflammation and neuronal damage. Display omitted •α-Syn is a ligand of RAGE receptor•vRAGE uses its positively charged surface to bind with acidic C terminus of α-syn•RAGE mediates the binding of α-syn amyloid fibrils to microglia•Neuroinflammation induced by α-syn fibrils can be mitigated by inhibition of RAGE Exploring the mechanism underlying the interplay between α-syn and neuroinflammation is important for PD treatment. Long et al. perform a series of biophysical and cellular experiments to demonstrate the structural basis of RAGE-α-syn interaction and the important role of RAGE in PD neuroinflammation. Blockage of the RAGE-α-syn interaction may alleviate α-syn-induced neuroinflammation in PD.