HAMLET Interacts with Lipid Membranes and Perturbs Their Structure and Integrity 英文参考文献.docVIP

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HAMLET Interacts with Lipid Membranes and Perturbs Their Structure and Integrity 英文参考文献.doc

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HAMLET Interacts with Lipid Membranes and Perturbs Their Structure and Integrity 英文参考文献

HAMLETInteractswithLipidMembranesandPerturbs TheirStructureandIntegrity Ann-KristinMossberg1.,MajaPuchades2.,?yvindHalskau3.,AnneBaumann3,IngelaLanekoff2 ,Yinxia Chao4,AuroraMartinez3,CatharinaSvanborg1,4*,RogerKarlsson2 1SectionofMicrobiology,Immunology,andGlycobiology,InstituteofLaboratoryMedicine,LundUniversity, Lund,Sweden, 2DepartmentofChemistry,Universityof Gothenburg,Go¨teborg,Sweden,3DepartmentofBiomedicine,UniversityofBergen,Bergen,Norway,4SingaporeImmunologyNetwork,A*STAR,Singapore,Singapore Abstract Background:Cellmembraneinteractionsrelyonlipidbilayerconstituentsandmoleculesinsertedwithinthemembrane, includingspecificreceptors.HAMLET(humana-lactalbuminmadelethaltotumorcells)isatumoricidalcomplexofpartially unfolded a-lactalbumin (HLA) and oleic acid that is internalized by tumor cells, suggesting that interactions with the phospholipid bilayer and/or specific receptors may be essential for the tumoricidal effect. This study examined whether HAMLETinteractswithartificialmembranesandaltersmembranestructure. Methodology/PrincipalFindings:WeshowbysurfaceplasmonresonancethatHAMLETbindswithhighaffinitytosurface adherent,unilamellarvesiclesoflipidswithvaryingacylchaincompositionandnetcharge.Fluorescenceimagingrevealed thatHAMLETaccumulatesinmembranesofvesiclesandperturbstheirstructure,resultinginincreasedmembranefluidity. Furthermore,HAMLETdisruptedmembraneintegrityatneutralpHandphysiologicalconditions,asshownbyfluorophore leakageexperiments.TheseeffectsdidnotoccurwitheithernativeHLAoraconstitutivelyunfoldedCys-AlaHLAmutant (rHLAall-Ala).HAMLETalsoboundtoplasmamembranevesiclesformedfromintacttumorcells,withaccumulationincertain membraneareas,butthecomplexwasnotinternalizedbythesevesiclesorbythesyntheticmembranevesicles. Conclusions/Significance:Theresultsillustratethedifferenceinmembraneaffinitybetweenthefattyacidboundandfatty acidfreeformsofpartiallyunfoldedHLAandsuggestthatHAMLETengagesmembranesbyamechanismrequiringboth theproteinandthefattyacid.Furth