title: Imaging the lipid-phase-dependent pore formation of equinatoxin II in droplet interface bilayers.
abstract: Using phase-separated droplet interface bilayers, we observe membrane binding and pore formation of a eukaryotic cytolysin, Equinatoxin II (EqtII). EqtII activity is known to depend on the presence of sphingomyelin in the target membrane and is enhanced by lipid phase separation. By imaging the ionic flux through individual pores in vitro, we observe that EqtII pores form predominantly within the liquid-disordered phase. We observe preferential binding of labeled EqtII at liquid-ordered/liquid-disordered domain boundaries before it accumulates in the liquid-disordered phase.
authors.affiliation: Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: N
authors.initials: N
authors.lastname: Rojko
authors.affiliation: Department of Chemistry, Oxford University, Oxford, UK.
authors.forename: B
authors.initials: B
authors.lastname: Cronin
authors.affiliation: Department of Chemistry, Oxford University, Oxford, UK.
authors.forename: J S H
authors.initials: JS
authors.lastname: Danial
authors.affiliation: Department of Chemistry, Oxford University, Oxford, UK.
authors.forename: M A B
authors.initials: MA
authors.lastname: Baker
authors.affiliation: Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia; National Institute of Chemistry, Ljubljana, Slovenia. Electronic address: gregor.anderluh@ki.si.
authors.forename: G
authors.initials: G
authors.lastname: Anderluh
authors.affiliation: Department of Chemistry, Oxford University, Oxford, UK. Electronic address: mark.wallace@chem.ox.ac.uk.
authors.forename: M I
authors.initials: MI
authors.lastname: Wallace
mesh_terms: D000818:Animals; D002645:Chickens; D003064:Cnidarian Venoms / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D066292:Lipid Droplets / Q000737:chemistry*; D057054:Molecular Imaging / Q000379:methods*; D016062:Porosity; D012756:Sheep
chemical_list: D003064:Cnidarian Venoms; D008051:Lipid Bilayers; C012307:equinatoxin
references.citation: Biochim Biophys Acta. 2011 Sep;1808(9):2275-88
references.pmid: 21621507
references.citation: J Biol Chem. 2004 Nov 5;279(45):46509-17
references.pmid: 15322132
references.citation: Structure. 2011 Feb 9;19(2):181-91
references.pmid: 21300287
references.citation: Trends Biochem Sci. 2008 Oct;33(10):482-90
references.pmid: 18778941
references.citation: Biochem J. 2000 Feb 15;346 Pt 1:223-32
references.pmid: 10657261
references.citation: Biochemistry. 1996 Nov 26;35(47):14947-57
references.pmid: 8942660
references.citation: Mol Biosyst. 2008 Dec;4(12):1191-208
references.pmid: 19396383
references.citation: Structure. 2003 Nov;11(11):1319-28
references.pmid: 14604522
references.citation: Nat Protoc. 2013 Jun;8(6):1048-57
references.pmid: 23640169
references.citation: Biochem Biophys Res Commun. 1998 Jan 6;242(1):187-90
references.pmid: 9439633
references.citation: FEBS J. 2006 Feb;273(4):863-71
references.pmid: 16441671
references.citation: Biochim Biophys Acta. 2012 Apr;1818(4):984-94
references.pmid: 21884678
references.citation: Biophys J. 2008 Jul;95(2):691-8
references.pmid: 18390598
references.citation: Biochim Biophys Acta. 2007 Sep;1768(9):2182-94
references.pmid: 17588529
references.citation: J Biol Chem. 2002 Nov 1;277(44):41916-24
references.pmid: 12198118
references.citation: J Biol Chem. 2003 Nov 14;278(46):45216-23
references.pmid: 12944411
references.citation: Cell Microbiol. 2011 Jul;13(7):1026-43
references.pmid: 21518219
references.citation: Biophys J. 2011 Dec 7;101(11):2679-83
references.pmid: 22261056
references.citation: Biochem Biophys Res Commun. 1996 Mar 18;220(2):437-42
references.pmid: 8645323
references.citation: Cell Calcium. 2005 Apr;37(4):283-99
references.pmid: 15755490
references.citation: FEMS Microbiol Immunol. 1992 Sep;5(1-3):121-9
references.pmid: 1384592
references.citation: Genes Cells. 2012 Aug;17(8):720-7
references.pmid: 22747662
references.citation: Cell Mol Life Sci. 2013 Jun;70(12):2083-98
references.pmid: 22983385
references.citation: Proc Natl Acad Sci U S A. 2011 Jan 25;108(4):1343-8
references.pmid: 21205902
references.citation: Biophys J. 2012 Feb 8;102(3):489-97
references.pmid: 22325271
references.citation: J Biol Chem. 2011 Oct 28;286(43):37768-77
references.pmid: 21885440
references.citation: Adv Exp Med Biol. 2010;677:106-15
references.pmid: 20687484
references.citation: J Am Chem Soc. 2009 Feb 11;131(5):1652-3
references.pmid: 19146373
references.citation: J Biol Chem. 2003 Jun 20;278(25):22678-85
references.pmid: 12676945
references.citation: J Membr Biol. 1993 Jan;131(1):11-22
references.pmid: 7679444
references.citation: J Mol Biol. 2005 Mar 18;347(1):27-39
references.pmid: 15733915
references.citation: J Biol Chem. 2013 Jan 11;288(2):1340-52
references.pmid: 23150664
references.citation: J Biol Chem. 2008 Jul 4;283(27):18665-77
references.pmid: 18442982
references.citation: J Am Chem Soc. 2006 Jan 11;128(1):192-201
references.pmid: 16390147
references.citation: Langmuir. 2011 Feb 15;27(4):1457-62
references.pmid: 21155607
references.citation: Prog Biophys Mol Biol. 2005 May;88(1):91-142
references.pmid: 15561302
references.citation: Biochim Biophys Acta. 2012 Jul;1818(7):1777-84
references.pmid: 22450237
references.citation: Toxicon. 2002 Feb;40(2):111-24
references.pmid: 11689232
references.citation: Eur J Biochem. 1999 Jul;263(1):128-36
references.pmid: 10429196
references.citation: J Biol Chem. 2010 Jul 16;285(29):22186-95
references.pmid: 20463009
references.citation: J Biol Chem. 2004 Aug 13;279(33):34209-16
references.pmid: 15175339
references.citation: Biochemistry. 1999 Nov 9;38(45):14999-5008
references.pmid: 10555982
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834
references.citation: Toxicon. 2009 Dec 15;54(8):1125-34
references.pmid: 19268680
references.citation: Proc Natl Acad Sci U S A. 2012 May 22;109(21):8097-102
references.pmid: 22562795
references.citation: J Am Chem Soc. 2011 Sep 21;133(37):14507-9
references.pmid: 21848341
references.citation: Nat Methods. 2008 Aug;5(8):695-702
references.pmid: 18641657
references.citation: Nat Rev Immunol. 2006 Dec;6(12):940-52
references.pmid: 17124515