Accumulated bending energy elicits neutral sphingomyelinase activity in human red blood cells

David J. López, Meritxell Egido-Gabas, Iván López-Montero, Jon V. Busto, Josefina Casas, Marie Garnier, Francisco Monroy, Banafshé Larijani, Félix M. Goñi, Alicia Alonso

Research output: Contribution to journalArticlepeer-review

Abstract

We propose that accumulated membrane bending energy elicits a neutral sphingomyelinase (SMase) activity in human erythrocytes. Membrane bending was achieved by osmotic or chemical processes, and SMase activity was assessed by quantitative thin-layer chromatography, high-performance liquid chromatography, and electrospray ionization-mass spectrometry. The activity induced by hypotonic stress in erythrocyte membranes had the pH dependence, ion dependence, and inhibitor sensitivity of mammalian neutral SMases. The activity caused a decrease in SM contents, with a minimum at 6 min after onset of the hypotonic conditions, and then the SM contents were recovered. We also elicited SMase activity by adding lysophosphatidylcholine externally or by generating it with phospholipase A2. The same effect was observed upon addition of chlorpromazine or sodium deoxycholate at concentrations below the critical micellar concentration, and even under hypertonic conditions. A unifying factor of the various agents that elicit this SMase activity is the accumulated membrane bending energy. Both hypo-and hypertonic conditions impose an increased curvature, whereas the addition of surfactants or phospholipase A2 activation increases the outer monolayer area, thus leading to an increased bending energy. The fact that this latent SMase activity is tightly coupled to the membrane bending properties suggests that it may be related to the general phenomenon of stress-induced ceramide synthesis and apoptosis.

Original languageEnglish
Pages (from-to)2077-2085
Number of pages9
JournalBiophysical Journal
Volume102
Issue number9
DOIs
Publication statusPublished - 1 May 2012

ASJC Scopus subject areas

  • Biophysics

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