Polyunsaturated fatty acids inhibit stimulated coupling between the ER Ca2 + sensor STIM1 and the Ca2 + channel protein Orai1 in a process that correlates with inhibition of stimulated STIM1 oligomerization

• The polyunsaturated fatty acid, linoleic acid, potently inhibits FcεRI signaling.
• Inhibition involves uncoupling of the ER sensor STIM1 from the Ca2 + channel Orai1.
• Results suggest that PUFAs regulate Ca2 + entry by perturbing ER membrane structure.

Polyunsaturated fatty acids (PUFAs) have been found to be effective inhibitors of cell signaling in numerous contexts, and we find that acute addition of micromolar PUFAs such as linoleic acid effectively inhibit of Ca2 + responses in mast cells stimulated by antigen-mediated crosslinking of FcεRI or by the SERCA pump inhibitor, thapsigargin. In contrast, the saturated fatty acid, stearic acid, with the same carbon chain length as linoleic acid does not inhibit these responses. Consistent with this inhibition of store-operated Ca2 + entry (SOCE), linoleic acid inhibits antigen-stimulated granule exocytosis to a similar extent. Using the fluorescently labeled plasma membrane Ca2 + channel protein, AcGFP–Orai1, together with the labeled ER Ca2 + sensor protein, STIM1–mRFP, we monitor stimulated coupling of these proteins that is essential for SOCE with a novel spectrofluorimetric resonance energy transfer method. We find effective inhibition of this stimulated coupling by linoleic acid that accounts for the inhibition of SOCE. Moreover, we find that linoleic acid induces some STIM1–STIM1 association, while inhibiting stimulated STIM1 oligomerization that precedes STIM1–Orai1 coupling. We hypothesize that linoleic acid and related PUFAs inhibit STIM1–Orai1 coupling by a mechanism that involves perturbation of ER membrane structure, possibly by disrupting electrostatic interactions important in STIM1 oligomerization. Thisarticle is part of a Special Issue entitled Tools to study lipid functions.