Departments of Molecular Medicine, The Scripps Research Institute, Scripps, Jupiter, United States; Scripps Research Skaggs Graduate School of Chemical and Biological Science, The Scripps Research Institute, Scripps,, Jupiter, United States
Mahmud Arif Pavel
Departments of Molecular Medicine, The Scripps Research Institute, Scripps, Jupiter, United States
Samuel S Hansen
Departments of Molecular Medicine, The Scripps Research Institute, Scripps, Jupiter, United States
Manasa Gudheti
Division of Endocrinology and Metabolism, Center for Brain Immunology and Glia, Department of Neuroscience, University of Virginia, Charlottesville, United States
Hao Wang
Departments of Molecular Medicine, The Scripps Research Institute, Scripps, Jupiter, United States; Scripps Research Skaggs Graduate School of Chemical and Biological Science, The Scripps Research Institute, Scripps,, Jupiter, United States
Departments of Molecular Medicine, The Scripps Research Institute, Scripps, Jupiter, United States; Scripps Research Skaggs Graduate School of Chemical and Biological Science, The Scripps Research Institute, Scripps,, Jupiter, United States
Keith R Murphy
Department of Neuroscience, The Scripps Research Institute, Scripps, Jupiter, United States; Center on Aging,The Scripps Research Institute, Scripps, Jupiter, United States
William Ja
Department of Neuroscience, The Scripps Research Institute, Scripps, Jupiter, United States; Center on Aging,The Scripps Research Institute, Scripps, Jupiter, United States
Heather A Ferris
Division of Endocrinology and Metabolism, Center for Brain Immunology and Glia, Department of Neuroscience, University of Virginia, Charlottesville, United States
Erik Jorgensen
Department of Biology, Howard Hughes Medical Institute, University of Utah, Salt Lake City, United States
Rapid conversion of force into a biological signal enables living cells to respond to mechanical forces in their environment. The force is believed to initially affect the plasma membrane and then alter the behavior of membrane proteins. Phospholipase D2 (PLD2) is a mechanosensitive enzyme that is regulated by a structured membrane-lipid site comprised of cholesterol and saturated ganglioside (GM1). Here we show stretch activation of TWIK-related K+ channel (TREK-1) is mechanically evoked by PLD2 and spatial patterning involving ordered GM1 and 4,5-bisphosphate (PIP2) clusters in mammalian cells. First, mechanical force deforms the ordered lipids, which disrupts the interaction of PLD2 with the GM1 lipids and allows a complex of TREK-1 and PLD2 to associate with PIP2 clusters. The association with PIP2 activates the enzyme, which produces the second messenger phosphatidic acid (PA) that gates the channel. Co-expression of catalytically inactive PLD2 inhibits TREK-1 stretch currents in a biological membrane. Cellular uptake of cholesterol inhibits TREK-1 currents in culture and depletion of cholesterol from astrocytes releases TREK-1 from GM1 lipids in mouse brain. Depletion of the PLD2 ortholog in flies results in hypersensitivity to mechanical force. We conclude PLD2 mechanosensitivity combines with TREK-1 ion permeability to elicit a mechanically evoked response.
