Zinc ions trigger the prion protein liquid-liquid phase separation.

Prion diseases are characterized by the misfolding and conversion of the monomeric prion protein (PrP) to a multimeric aggregated pathogenic form, known as PrPSc. We and others have recently shown that biomolecular condensates formed via liquid-liquid phase separation of PrP can undergo maturation to solid-like species that resemble pathological aggregates, and this process is modulated by DNA, RNA, and oxidative conditions. Conversely, the most well-studied ligand of PrP, copper ions, induce liquid-like condensates of PrP that accumulate Cu2+in vitro, and live PrPC-expressing cells show condensation at the cell surface as triggered by physiologically relevant conditions of Cu2+ and protein concentrations. Since PrP can also bind to Zn2+ through its intrinsically disordered N-terminal domain, though with different affinities and binding modes than Cu2+, we hypothesized that Zn2+ could modulate PrP phase separation differently from copper ions. Using an appropriate buffer with negligible metal ion binding, as well as relevant pH, ionic strength, molecular crowding, and Zn2+ concentrations, we show that recombinant PrP undergoes phase separation with Zn2+. Furthermore, we show that metal ion-induced condensation of PrP is dependent on the N-terminal domain (residues 23-90). In vitro Fluorescence Recovery After Photobleaching (FRAP) experiments and thioflavin T aggregation kinetics support key differences in the molecular properties of PrP:Zn2+versus PrP:Cu2+ phase separated states. FRAP analysis indicated that both Cu2+ and Zn2+ promote liquid-like PrP condensates; however, PrP:Zn2+condensates exhibit a faster recovery. Cu2+ pronouncedly inhibits seed-induced PrP misfolding, whereas Zn2+ provides a milder delay in PrP aggregation. Our findings provide insights on Zn2+-induced phase separation of PrP, supporting a variety of previously proposed functions of PrP in metal sequestering and uptake, processes that could be effectively regulated through biomolecular condensation. Copyright © 2025 Elsevier Inc. All rights reserved.

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Mariana Juliani do Amaral, Letícia Soares de Oliveira, Yraima Cordeiro. Zinc ions trigger the prion protein liquid-liquid phase separation. Biochemical and biophysical research communications. 2025 Mar 19;753:151489

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PMID: 39983547

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