title: Cholesterol modulates Orai1 channel function.
abstract: STIM1 (stromal interaction molecule 1) and Orai proteins are the essential components of Ca(2+) release-activated Ca(2+) (CRAC) channels. We focused on the role of cholesterol in the regulation of STIM1-mediated Orai1 currents. Chemically induced cholesterol depletion enhanced store-operated Ca(2+) entry (SOCE) and Orai1 currents. Furthermore, cholesterol depletion in mucosal-type mast cells augmented endogenous CRAC currents, which were associated with increased degranulation, a process that requires calcium influx. Single point mutations in the Orai1 amino terminus that would be expected to abolish cholesterol binding enhanced SOCE to a similar extent as did cholesterol depletion. The increase in Orai1 activity in cells expressing these cholesterol-binding-deficient mutants occurred without affecting the amount in the plasma membrane or the coupling of STIM1 to Orai1. We detected cholesterol binding to an Orai1 amino-terminal fragment in vitro and to full-length Orai1 in cells. Thus, our data showed that Orai1 senses the amount of cholesterol in the plasma membrane and that the interaction of Orai1 with cholesterol inhibits its activity, thereby limiting SOCE.
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria. isabella.derler@jku.at christoph.romanin@jku.at.
authors.forename: Isabella
authors.initials: I
authors.lastname: Derler
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Isaac
authors.initials: I
authors.lastname: Jardin
authors.affiliation: Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario N6A 5C1, Canada.
authors.forename: Peter B
authors.initials: PB
authors.lastname: Stathopulos
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Muik
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Marc
authors.initials: M
authors.lastname: Fahrner
authors.affiliation: Center for Nanobiology and Structural Biology, Institute of Microbiology, Academy of Sciences of the Czech Republic, CZ-373 33 Nové Hrady, Czech Republic.
authors.forename: Vasilina
authors.initials: V
authors.lastname: Zayats
authors.affiliation: Center for Nanobiology and Structural Biology, Institute of Microbiology, Academy of Sciences of the Czech Republic, CZ-373 33 Nové Hrady, Czech Republic.
authors.forename: Saurabh K
authors.initials: SK
authors.lastname: Pandey
authors.affiliation: Institute of Biophysics, Medical University of Graz, Harrachgasse 21/4, 8010 Graz, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Poteser
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Barbara
authors.initials: B
authors.lastname: Lackner
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Marketa
authors.initials: M
authors.lastname: Absolonova
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria.
authors.forename: Rainer
authors.initials: R
authors.lastname: Schindl
authors.affiliation: Institute of Biophysics, Medical University of Graz, Harrachgasse 21/4, 8010 Graz, Austria.
authors.forename: Klaus
authors.initials: K
authors.lastname: Groschner
authors.affiliation: Center for Nanobiology and Structural Biology, Institute of Microbiology, Academy of Sciences of the Czech Republic, CZ-373 33 Nové Hrady, Czech Republic.
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Ettrich
authors.affiliation: Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 1L7, Canada.
authors.forename: Mitsu
authors.initials: M
authors.lastname: Ikura
authors.affiliation: Institute of Biophysics, Johannes Kepler University of Linz, Gruberstrasse 40, 4020 Linz, Austria. isabella.derler@jku.at christoph.romanin@jku.at.
authors.forename: Christoph
authors.initials: C
authors.lastname: Romanin
mesh_terms: D019921:Biotinylation; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D002789:Cholesterol Oxidase / Q000378:metabolism; D002942:Circular Dichroism; D055724:Electrophysiological Phenomena; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006632:Histamine / Q000378:metabolism; D006801:Humans; D008407:Mast Cells / Q000378:metabolism; D009154:Mutation; D000071740:ORAI1 Protein; D010455:Peptides / Q000378:metabolism; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence
chemical_list: D015220:Calcium Channels; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; D010455:Peptides; D006632:Histamine; D002784:Cholesterol; D002789:Cholesterol Oxidase; D002118:Calcium
references.citation: Am J Physiol Cell Physiol. 2010 May;298(5):C993-1005
references.pmid: 20107038
references.citation: J Biol Chem. 2011 Jul 15;286(28):24966-76
references.pmid: 21555515
references.citation: Adv Exp Med Biol. 2012;740:383-410
references.pmid: 22453951
references.citation: Biochemistry. 1995 Oct 24;34(42):13784-93
references.pmid: 7577971
references.citation: Biochem J. 2009 Dec 14;425(1):159-68
references.pmid: 19843011
references.citation: Proc Natl Acad Sci U S A. 2012 May 29;109 (22):8682-7
references.pmid: 22586105
references.citation: Am J Physiol Cell Physiol. 2008 Jun;294(6):C1323-31
references.pmid: 18400989
references.citation: Cell Calcium. 2015 Oct;58(4):342-8
references.pmid: 25843208
references.citation: Curr Biol. 2005 Jul 12;15(13):1235-41
references.pmid: 16005298
references.citation: Nat Cell Biol. 2010 May;12 (5):436-46
references.pmid: 20418871
references.citation: Biophys J. 2012 Nov 7;103(9):1899-908
references.pmid: 23199918
references.citation: Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3202-6
references.pmid: 19221033
references.citation: Proc Natl Acad Sci U S A. 2009 Sep 8;106(36):15495-500
references.pmid: 19706428
references.citation: J Biol Chem. 2009 Mar 27;284(13):8421-6
references.pmid: 19189966
references.citation: Biophys J. 2004 Oct;87(4):2419-32
references.pmid: 15454440
references.citation: Endocrinology. 1998 Dec;139(12 ):4991-7
references.pmid: 9832438
references.citation: J Biol Chem. 2013 Oct 25;288(43):31154-64
references.pmid: 24019518
references.citation: J Biol Chem. 2012 Oct 12;287(42):35612-20
references.pmid: 22932896
references.citation: Biochemistry. 2010 Oct 26;49(42):9027-31
references.pmid: 20863067
references.citation: Cell. 2012 Apr 13;149(2):425-38
references.pmid: 22464749
references.citation: Biochim Biophys Acta. 2011 Mar;1813(3):431-7
references.pmid: 21255618
references.citation: Traffic. 2008 Nov;9(11):1839-49
references.pmid: 18647169
references.citation: J Biol Chem. 2009 Jun 5;284(23 ):15903-15
references.pmid: 19366689
references.citation: Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6295-9
references.pmid: 8392195
references.citation: Adv Second Messenger Phosphoprotein Res. 1999;33:279-307
references.pmid: 10218123
references.citation: Cell. 2009 Mar 6;136(5):876-90
references.pmid: 19249086
references.citation: Biophys J. 2011 Oct 19;101(8):1959-67
references.pmid: 22004750
references.citation: Biochem Biophys Res Commun. 2015 Apr 24;460(1):40-9
references.pmid: 25998732
references.citation: Nat Neurosci. 2006 Sep;9(9):1108-16
references.pmid: 16921373
references.citation: Nature. 2005 Oct 6;437(7060):902-5
references.pmid: 16208375
references.citation: J Physiol. 2012 Sep 1;590(17):4169-77
references.pmid: 22586216
references.citation: Biophys J. 2003 Jun;84(6):3992-4010
references.pmid: 12770904
references.citation: J Exp Med. 2005 Sep 5;202(5):651-62
references.pmid: 16147976
references.citation: Front Biosci (Landmark Ed). 2012 Jan 01;17:1304-22
references.pmid: 22201805
references.citation: Sci Signal. 2013 Mar 19;6(267):pe9
references.pmid: 23512988
references.citation: J Biol Chem. 2007 Mar 9;282(10):7276-86
references.pmid: 17209047
references.citation: FEBS Lett. 1992 Sep 28;310(2):123-8
references.pmid: 1397260
references.citation: Nat Rev Mol Cell Biol. 2003 Jul;4(7):517-29
references.pmid: 12838335
references.citation: J Biol Chem. 2008 Mar 21;283(12 ):8014-22
references.pmid: 18187424
references.citation: J Cell Sci. 2011 Aug 1;124(Pt 15):2602-10
references.pmid: 21750194
references.citation: Nature. 2006 May 11;441(7090):179-85
references.pmid: 16582901
references.citation: J Biol Chem. 1985 Mar 25;260(6):3440-50
references.pmid: 3838314
references.citation: Biochim Biophys Acta. 2008 Jul-Aug;1778(7-8):1576-82
references.pmid: 18423371
references.citation: J Biol Chem. 2012 Dec 14;287(51):43030-41
references.pmid: 23109337
references.citation: Biophys J. 2001 Oct;81(4):2395-402
references.pmid: 11566809
references.citation: J Phys Chem B. 2014 Nov 20;118(46):13169-74
references.pmid: 25347282
references.citation: Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):1267-72
references.pmid: 11158628
references.citation: Methods Mol Biol. 2015;1232:91-102
references.pmid: 25331130
references.citation: Biophys J. 2015 Jul 7;109(1):85-94
references.pmid: 26153705
references.citation: Biochim Biophys Acta. 2010 Sep;1803(9):1083-93
references.pmid: 20600358
references.citation: Mol Biol Cell. 2009 Jan;20(1):389-99
references.pmid: 18987344
references.citation: EMBO J. 2011 May 4;30(9):1678-89
references.pmid: 21427704
references.citation: Channels (Austin). 2008 Nov-Dec;2(6):401-3
references.pmid: 18843204
references.citation: Gen Physiol Biophys. 2009 Jun;28(2):117-25
references.pmid: 19592708
references.citation: Biochemistry. 2003 Oct 14;42(40):11808-14
references.pmid: 14529292
references.citation: Mol Biol Cell. 2008 Jul;19(7):2802-17
references.pmid: 18448669
references.citation: Cancer Res. 2013 Aug 1;73(15):4852-61
references.pmid: 23774210
references.citation: J Biol Chem. 2013 Oct 4;288(40):29025-34
references.pmid: 23943619
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):9357-62
references.pmid: 16751269
references.citation: Science. 2006 May 26;312(5777):1220-3
references.pmid: 16645049
references.citation: Science. 2012 Dec 7;338(6112):1308-13
references.pmid: 23180775
references.citation: Biosci Rep. 2007 Dec;27(6):335-58
references.pmid: 17668316
references.citation: J Physiol. 2006 Nov 15;577(Pt 1):31-44
references.pmid: 16959851
references.citation: Nat Struct Mol Biol. 2010 Jan;17 (1):112-6
references.pmid: 20037597
references.citation: J Exp Med. 2006 May 15;203(5):1161-71
references.pmid: 16618793