Acylcarnitines at the Membrane Surface: Insertion Parameters for a Mitochondrial Leaflet Model.

Excessive accumulation of acylcarnitines (ACs), often caused by metabolic disorders, has been associated with obesity, arrhythmias, cardiac ischemia, insulin resistance, etc. Mechanisms whereby elevated ACs might contribute to pathophysiological effects remain largely unexplored. We have aimed to gain insight into AC interactions with the mitochondrial inner membrane. To model its outer leaflet, Langmuir monolayers and cushioned supported bilayers were employed. Their interactions with ACs were monitored with epifluorescence microscopy, which revealed a local leaflet expansion upon exposure to elevated concentrations of a long-chain AC, plausibly caused by its insertion. To assess the AC insertion parameters, constant-pressure insertion assays were performed. A value of 21 ± 3 Å2 was obtained for the AC insertion area, which is roughly the same as the cross-sectional area of an acyl chain. By contrast, the carnitine moiety was found to require an area of 37 ± 3 Å2. The AC insertion has thus been concluded to involve solely the AC acyl chain. This mode of insertion implies that the carnitine moiety, with its nontitratable positive charge, is left dangling at the membrane surface, which is likely to alter the surface electrostatics of the outer leaflet. The extrapolation of these findings has enabled us to hypothesize that, by altering the morphology and surface electrostatics of the outer leaflet, the insertion of ACs, in particular their long-chain counterparts, may trigger a nonspecific activation of signaling pathways in the inner mitochondrial membrane, thereby modulating its function and potentially leading to pathophysiological responses.