Use of novel synthetic analogs to decipher the interactome of phosphatidic acid species in neurosecretory cells.

Fiche publication

Date publication

janvier 2026

Journal

Methods in enzymology

Auteurs

Membres identifiés du Cancéropôle Est :
Dr GASMAN Stéphane , Dr VITALE Nicolas , Dr ORY Stéphane

Tous les auteurs :
Wolf A, Tanguy E, Decraene C, Vertueux S, Strub JM, Ory S, Balieu S, Renard PY, Gasman S, Vitale N

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/41895912

Résumé

Understanding how phospholipids orchestrate cellular signaling requires tools that preserve their native complexity to reveal their modular function. Here, we introduce a versatile chemical approach to map the phosphatidic acid (PA) interactome directly within membranes of living cells. Using synthetic PA analogues, modified on the glycerol backbone with a bioorthogonal azide group, we generated functional probes that retain natural headgroup and acyl-chain composition and properties. These analogues can be coupled to photoactivatable crosslinkers, allowing covalent capture of transient PA-protein partners upon UV illumination. We present here approaches to validate their integration and functionality through recruitment assays using PA-binding sensor and ERK1/2 phosphorylation readouts. Applying this framework in neurosecretory PC12 cells, we identified specific PA-binding partners across distinct secretory states, revealing both acyl chain-dependent and activity-specific interaction patterns. This methodology bridges lipid chemistry and cell biology by enabling dynamic, species-specific exploration of phospholipid signaling networks. Beyond neurosecretion, the approach provides a broadly applicable platform for dissecting lipid-mediated processes across diverse physiological and pathological contexts, from membrane trafficking to disease-associated lipid signaling.