RRC ID 67361
Author Suzuki J, Umeda M, Sims PJ, Nagata S.
Title Calcium-dependent phospholipid scrambling by TMEM16F.
Journal Nature
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.
Volume 468(7325)
Pages 834-8
Published 2010-12-9
DOI 10.1038/nature09583
PII nature09583
PMID 21107324
MeSH 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
IF 42.779
DNA material pMXs-puro-mTMEM16A (RDB18742) pMXs-puro-mTMEM16B (RDB18743) pMXs-puro-mTMEM16C (RDB18744) pMXs-puro-mTMEM16D (RDB18745) pMXs-puro-mTMEM16E (RDB18746) pMXs-puro-mTMEM16F (RDB18747) pMXs-puro-mTMEM16G (RDB18748) pMXs-puro-mTMEM16H (RDB18749) pMXs-puro-mTMEM16J (RDB18750) pMXs-puro-mTMEM16F-long (RDB18751) pMXs-puro-mTMEM16F-D409G (RDB18752) pMXs-puro-mTMEM16F-D409G long (RDB18753) pMXs-puro-mTMEM16F-R211L (RDB18754) pMXs-puro-mTMEM16F-D409G/R211L (RDB18755) pEF-mTMEM16A (RDB18756) pEF-mTMEM16B (RDB18757) pEF-mTMEM16C (RDB18758) pEF-mTMEM16D (RDB18759) pEF-mTMEM16E (RDB18760) pEF-mTMEM16F (RDB18761) pEF-mTMEM16G (RDB18762) pEF-mTMEM16H (RDB18763) pEF-mTMEM16J (RDB18764) pEF-mTMEM16K (RDB18765)