Petasin at the Membrane: Fine-Tuning Lipid Organization and Interfacial Electrostatics in Mast Cell Plasma Membrane Models

Petasin, a lipophilic sesquiterpene ester isolated from Petasites hybridus, exhibits well-documented anti-inflammatory and anti-allergic activities, although its molecular mechanisms of action remain incompletely understood. Here, we investigated its direct effects on membrane organization using biomimetic large unilamellar vesicles (LUVs) modeling resting and activated mast cell plasma membranes. The models comprised phosphatidylcholines (EggPC, PAPC), sphingomyelin (SM), ganglioside GD3, and cholesterol (Chol), with phosphatidylserine (PS) additionally incorporated into the activated membrane model. Simplified liquid-disordered (Ld) and liquid-ordered (Lo) lipid systems were used to resolve phase-dependent effects. Membrane properties were assessed by Laurdan generalized polarization, DPH fluorescence anisotropy, DPH-TEMPO fluorescence quenching, di-8-ANEPPS dipole potential measurements, and dynamic light scattering (DLS). Petasin exerted phase- and composition-dependent effects on membrane organization. It increased lipid order in relatively disordered membrane environments, while producing only minor changes in highly ordered Lo membranes, revealing a clear dependence of its effects on the initial lipid phase state. In heterogeneous mast cell membrane models exhibiting Lo/Ld phase coexistence, DPH-TEMPO quenching indicated a concentration-dependent increase in the estimated size of ordered lipid nanodomains. In parallel, petasin decreased membrane dipole potential in a concentration-dependent manner, demonstrating modulation of membrane interfacial electrostatic properties. These findings indicate coordinated modulation of lipid packing, lateral membrane organization, and interfacial electrostatics, with the membrane response influenced by lipid composition. DLS measurements showed no evidence of vesicle aggregation or major alterations in vesicle size distribution. Overall, these findings identify petasin as a membrane-active modulator capable of fine-tuning lipid order, lateral membrane organization, and interfacial electrostatics in a phase- and composition-dependent manner. Given the importance of membrane organization and lipid raft dynamics in FcεRI clustering and mast cell activation, these membrane effects may represent an additional biophysical mechanism of the anti-inflammatory and anti-allergic actions of petasin. More broadly, our findings highlight membrane physicochemical organization as a relevant factor in the regulation of mast cell function and provide a basis for further investigation of membrane-mediated mechanisms in allergic and mast cell-associated inflammatory responses.