Calcium-dependent Phospholipid Scrambling by TMEM16F

Nature. 2010 Dec 9;468(7325):834-8. doi: 10.1038/nature09583. Epub 2010 Nov 24.

Abstract

In all animal cells, phospholipids are asymmetrically distributed between the outer and inner leaflets of the plasma membrane. This asymmetrical phospholipid distribution is disrupted in various biological systems. For example, when blood platelets are activated, they expose phosphatidylserine (PtdSer) to trigger the clotting system. The PtdSer exposure is believed to be mediated by Ca(2+)-dependent phospholipid scramblases that transport phospholipids bidirectionally, but its molecular mechanism is still unknown. Here we show that TMEM16F (transmembrane protein 16F) is an essential component for the Ca(2+)-dependent exposure of PtdSer on the cell surface. When a mouse B-cell line, Ba/F3, was treated with a Ca(2+) ionophore under low-Ca(2+) conditions, it reversibly exposed PtdSer. Using this property, we established a Ba/F3 subline that strongly exposed PtdSer by repetitive fluorescence-activated cell sorting. A complementary DNA library was constructed from the subline, and a cDNA that caused Ba/F3 to expose PtdSer spontaneously was identified by expression cloning. The cDNA encoded a constitutively active mutant of TMEM16F, a protein with eight transmembrane segments. Wild-type TMEM16F was localized on the plasma membrane and conferred Ca(2+)-dependent scrambling of phospholipids. A patient with Scott syndrome, which results from a defect in phospholipid scrambling activity, was found to carry a mutation at a splice-acceptor site of the gene encoding TMEM16F, causing the premature termination of the protein.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anoctamins
  • B-Lymphocytes / cytology
  • B-Lymphocytes / drug effects
  • Calcium / antagonists & inhibitors
  • Calcium / metabolism*
  • Calcium / pharmacology
  • Cell Line
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Flow Cytometry
  • Gene Library
  • Humans
  • Ionophores / pharmacology
  • Mice
  • Mutant Proteins / chemistry
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Phosphatidylserines / metabolism
  • Phospholipid Transfer Proteins / chemistry
  • Phospholipid Transfer Proteins / genetics
  • Phospholipid Transfer Proteins / metabolism*
  • Phospholipids / metabolism*
  • RNA Splice Sites / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Syndrome

Substances

  • Ano6 protein, human
  • Anoctamins
  • DNA, Complementary
  • Ionophores
  • Mutant Proteins
  • Phosphatidylserines
  • Phospholipid Transfer Proteins
  • Phospholipids
  • RNA Splice Sites
  • Calcium