Regulation of TMEM16A/ANO1 and TMEM16F/ANO6 ion currents and phospholipid scrambling by Ca2+ and plasma membrane lipid.

Abstract

KEY POINTS:

TMEM16 proteins can operate as Ca2+ -activated Cl- channels or scramble membrane phospholipids, which are both highly relevant mechanisms during disease. Overexpression of TMEM16A and TMEM16F were found to be partially active at 37°C and at resting intracellular Ca2+ concentrations. We show that TMEM16 Cl- currents and phospholipid scrambling can be activated by modification of plasma membrane phospholipids, through reactive oxygen species and phospholipase A2. While phospholipids and Cl- ions are likely to use the same pore within TMEM16F, TMEM16A only conducts Cl- ions. Lipid regulation of TMEM16 proteins is highly relevant during inflammation and regulated cell death such as apoptosis and ferroptosis.

ABSTRACT:

TMEM16/anoctamin (ANO) proteins form Ca2+ -activated ion channels or phospholipid scramblases. We found that both TMEM16A/ANO1 and TMEM16F/ANO6 produced Cl- currents when activated by intracellular Ca2+ , but only TMEM16F was able to expose phosphatidylserine to the outer leaflet of the plasma membrane. Mutations within TMEM16F or TMEM16A/F chimeras similarly changed Cl- currents and phospholipid scrambling, suggesting the same intramolecular pathway for Cl- and phospholipids. When overexpressed, TMEM16A and TMEM16F produced spontaneous Cl- currents at 37°C even at resting intracellular Ca2+ levels, which was abolished by inhibition of phospholipase A2 (PLA2 ). Connversely, activation of PLA2 or application of active PLA2 , as well as lipid peroxidation induced by reactive oxygen species (ROS) using staurosporine or tert-butyl hydroperoxide, enhanced ion currents by TMEM16A/F and in addition activated phospholipid scrambling by TMEM16F. Thus, TMEM16 proteins are activated by an increase in intracellular Ca2+ , or independent of intracellular Ca2+ , by modifications occurring in plasma and intracellular membrane phospholipids. These results may help to explain why regions distant to the TMEM16 pore and the Ca2+ binding sites control Cl- currents and phospholipid scrambling. Regulation of TMEM16 proteins through modification of membrane phospholipids occurs during regulated cell death such as apoptosis and ferroptosis. It contributes to inflammatory and nerve injury-induced hypersensitivity and generation of pain and therefore provides a regulatory mechanism that is particularly relevant during disease.

• Authors: Rainer Schreiber, Jiraporn Ousingsawat, Podchanart Wanitchakool, Lalida Sirianant, Roberta Benedetto, Karina Reiss, Karl Kunzelmann
• Journal: The Journal of physiology (J Physiol) Vol. 596 Issue 2 Pg. 217-229 (01 15 2018) ISSN: 1469-7793 [Electronic] England
• PMID: 29134661 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2017 The Authors. The Journal of Physiology © 2017 The Physiological Society.

Chemical References

• ANO1 protein, human
• ANO6 protein, human
• Anoctamin-1
• Anoctamins
• Calcium-Regulating Hormones and Agents
• Membrane Lipids
• Neoplasm Proteins
• Phospholipid Transfer Proteins
• Phospholipids
• Phospholipases A2
• Calcium

Topics

• Anoctamin-1 (genetics, metabolism)
• Anoctamins (genetics, metabolism)
• Apoptosis
• Calcium (pharmacology)
• Calcium-Regulating Hormones and Agents (pharmacology)
• Cell Membrane (metabolism)
• HEK293 Cells
• Humans
• Ion Transport
• Membrane Lipids (metabolism)
• Neoplasm Proteins (genetics, metabolism)
• Phospholipases A2 (metabolism)
• Phospholipid Transfer Proteins (genetics, metabolism)
• Phospholipids (metabolism)
• Signal Transduction