E ff ect of Protein Corona on Nanoparticle − Lipid Membrane Binding The Binding Strength and Dynamics - Lee - 2021 - Unknown

《E ff ect of Protein Corona on Nanoparticle − Lipid Membrane Binding The Binding Strength and Dynamics - Lee - 2021 - Unknown》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

pubs.acs.org/LangmuirArticleEffectofProteinCoronaonNanoparticle−LipidMembraneBinding:TheBindingStrengthandDynamicsHwankyuLee*CiteThis:Langmuir2021,37,3751−3760ReadOnlineACCESSMetrics&MoreArticleRecommendationsABSTRACT:All-atommoleculardynamicssimulationsofthe10nm-sizedanionicpolystyrene(PS)particlecomplexedwithplasmaproteins(humanserumalbumin,immunoglobulingamma-1chain-C,andapolipoproteinA-I)adsorbedontolipidbilayers[asymmetricallycomposedofextracellular(zwitterionic)andcytosolic(anionic)leaflets]areperformed.Freeenergiescalculatedfromumbrellasamplingsimulationsshowthatproteinsontheparticlemoreweaklybindtothezwitterionicleafletthandobareparticles,inagreementwithexperimentsshowingthesuppressionoftheparticle−bilayerbindingbyproteincorona.Proteinsontheparticleinteractmorestronglywiththeanionicleafletthanwiththezwitterionicleafletbecauseofchargeinteractionsbetweencationicproteinresiduesandanioniclipidheadgroups,toanextentdependentonvariousplasmaproteins.Inparticular,hydrogenbondsbetweenproteinsandzwitterionicleafletsrestrictthemotionoflipidsandthusreducethelateraldynamicsofbilayers,whilethetightbindingbetweenproteinsandanionicleafletsdisruptsthehelicalstructureofproteinsanddisorderslipids,leadingtoanincreaseinthelateraldynamicsofbilayers.Thesefindingshelpexplaintheexperimentalobservationregardingthefactthatthebilayerdynamicsdecreaseswheninteractingwithproteincoronaandsuggestthattheeffectofproteincoronaonthebindingstrengthandbilayerdynamicsdependsonproteintypesandbilayercharges.DownloadedviaUNIVOFNEWMEXICOonMay16,2021at06:31:31(UTC).■INTRODUCTIONtoanionicgoldnanoparticles,whichinducestheirinteractions14Seehttps://pubs.acs.org/sharingguidelinesforoptionsonhowtolegitimatelysharepublishedarticles.withintegrinreceptors.Tenzeretal.alsofoundthatcoronaNanoparticleshavebeenwidelyappliedforantitumor1−6formationpromotescellularuptakeofsilicaandpolystyrenetherapeuticsanddrug-deliveryapplications.Whendrug-15(PS)nanoparticles.Theseresultsindicatethatproteincoronaencapsulatingnanoparticlesflowthroughthebloodstream,theircanpromotecellularuptakeandinternalizationmostlyviacellsurfacesarerapidlycoatedbyplasmaproteins,leadingtothesurfacereceptors,whilemanyotherexperimentshavealsoformationofproteinlayersontheparticlesurface,calledprotein7,8showntheoppositetendency.Lesniaketal.observedlesscorona.Proteincoronaconsistsoftheinnerproteinlayeradhesionandinternalizationofsilicananoparticlesintocellstronglyboundtotheparticlesurface(hardcorona)andthe16membranesinthepresenceofproteincorona.Salvatietal.outerproteinlayerweaklyboundtotheboundaryoftheinner7,9showedthatproteincoronastericallyblocksthespecificbindingproteinlayer(softcorona).Therefore,whennanoparticlesarebetweentransferrin-conjugatednanoparticlesandtransferrintargetedtospecificcells,cellmembranesindeedrecognizeproteinlayersonparticlesurfacesratherthanbareparticlesurfaces,whichhasmotivatedmanyexperimentalandReceived:January26,2021theoreticalstudiesontheeffectofproteincoronaontheRevised:March1,2021interactionsbetweennanoparticlesandcellmembranes.10−12Published:March19,2021Experimentshaveshownthatcoronaformationinfluences13cellularuptakeandinternalizationofnanoparticles.Forinstance,Dengetal.observedunfoldingoffibrinogenbound©2021AmericanChemicalSocietyhttps://doi.org/10.1021/acs.langmuir.1c002493751Langmuir2021,37,3751−3760

1Langmuirpubs.acs.org/LangmuirArticle17receptorsoncells.Wangetal.foundthereducedcytotoxicitycomplexandbilayersurfaces,favorablycomparedwithexperi-18ofgoldnanoparticlescoatedwithproteincorona,andYanetal.mentswithmodelmembranes.Inparticular,weanalyzetheshowedthatcellularinternalizationofprotein-coatedparticleseffectoftheprotein−bilayerbindingontheproteinstructuredecreasesinhumanmonocyticcellsbutincreasesindiffer-andbilayerdynamics,whicharerationalizedbyhydrogen-bond19entiatedmacrophage-likecellsviaascavengerreceptor.interactionsbetweenchargedproteinresiduesandlipidKokkinopoulouetal.andSimonetal.,respectively,observedheadgroups.WewillshowthattheseresultshelpexplaininthereducedcellularuptakeforPSparticlescoatedwithsoftdetailtheexperimentalobservationsregardingthesuppression20coronaorapolipoproteinA-I(APO)andforthosecoatedwithoftheparticle−bilayerbindingbyproteincoronaandthe21plasmaproteinswithoutimmunoglobulingamma-1(IgG).Toreducedlateraldynamicsofbilayersinteractingwithproteininterpretthesedifferenteffectsofproteincoronaonthecorona.particle−cellinteraction,nonspecificinteractionsbetweenproteincoronaandsyntheticmodelmembraneshavebeen■METHODSsystematicallystudiedthroughinvitroexperiments.Montisetal.AllsimulationsandanalyseswereperformedusingtheGROMACS-showedthatthebindingbetweenproteincorona-coatedgold41−432018.6simulationpackagewiththeoptimizedpotentialforliquidnanoparticlesandmembranesurfacesinducestheraft-likesimulations(OPLS)all-atomforcefield(FF)andtheTIP4Pwater(ordered)phase,leadingtoadecreaseinthelateraldynamicsofmodel.44,45PotentialparametersforPSweretakenfromtheOPLSmembranes.22DiSilvioetal.showedthathardcoronamore46benzeneFFdevelopedbySeveranceandJorgensen,whichweaklyinteractswithmembranesthandobareanionicPSsuccessfullyreproducestheexperimentaldensity,conformation,and47particles,whilesoftcoronasignificantlydisruptsmembranesdynamicsofPSchains.Wepreviouslyequilibratedthe10nm-sizedthandofreeplasmaproteins.23Recently,Montisetal.24andanionicPSparticlecomposedof196PSchains(20styrenemonomersWangetal.11showedthatproteincoronaweakenselectrostaticperchain)modifiedbyaddingacarboxylgrouptothechainterminal(ainteractionsbetweennanoparticlesandmembranesurfacesandnetchargeof−196perparticle)andthensimulatedthePSparticleboundtovariousplasmaproteinssuchasSA,IgG,andAPOthatwerethussuppressestheadsorptionofnanoparticlestoboth48downloadedfromtheProteinDataBank(PDBcode:1AO6,macrophageplasmaandsyntheticmodelmembranes,confirm-4950351HZH,and3K2S,respectively).TheseequilibratedPSingthesuccessfulapplicationofmodelmembranesfortheparticle−proteincomplexeswereusedasstartingstatesinthiswork.proteincorona−cellinteraction.For1-palmitoyl-2-oleoyl-glycero-3-phosphocholine(POPC),1-palmi-Tounderstandandcomplementtheseexperimentalresultsattoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine(POPE),1-palmito-nearlytheatomicscale,moleculardynamics(MD)simulationsyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS),sphingomyelinhavebeenperformed.Mostsimulationstudieshavefocusedon(SM,d18:1/24:0),andcholesterol(CHOL),weusedpotentialthedynamicsandstructuralchangesofadsorbedproteinsandparameterscompatiblewiththeOPLSFFdevelopedbyRoǵetal.,theirinteractionswithdifferentlysized,shaped,andchargedwhichsuccessfullyreproducesexperimentalvaluesofareaperlipid,51−5425−34orderparameter,andlateraldynamicsoflipidbilayers.Roǵetal.nanoparticles.Recently,weperformedall-atomsimula-alsoconstructedamodelmembranecomposedofPOPC(9mol%),tionsoffivemajorplasmaproteinsadsorbedontocationic,POPE(18mol%),POPS(36mol%),andCHOL(37mol%)intheanionic,andhydrophobicPSparticles,showingthehigheranionicinner(cytosolic)leafletandPOPC(23mol%),SM(30molmobilityofproteinsintheouterproteinlayerthanintheinner%),andCHOL(47mol%)inthezwitterionicouter(extracellular)35proteinlayer,whichhelpstoexplainexperimentalobserva-leaflet,mimickingtheasymmetriccompositionofeukaryoticplasma55tions(i.e.,theVromaneffect)regardingcompetitiveadsorptionmembranes,whichweusedforthiswork.anddesorptionofplasmaproteinsonparticlesurfacesandtheAtemperatureof310Kandapressureof1barweremaintainedby56differenceinthebindingstrengthofproteinsininnerandouterapplyingavelocity-rescalethermostatandParrinello−Rahman57layers.36,37Besidestheseprotein−particleinteractions,theeffectbarostatinanNPxyPzTensemblewithsemi-isotropicpressurecoupling.Arealspacecutoffof1nmwasappliedforLennard-Jonesandofproteincoronaontheparticle−membraneinteractionhas3839electrostaticforceswiththeinclusionofparticlemeshEwaldbeenalsostudied.Huetal.andDingandMaperformedsummationforlong-rangeelectrostatics.58TheLINCSalgorithmwascoarse-grained(CG)simulationsofprotein-coatednano-usedtoconstraintthebondlengths.59,60Simulationswereperformedparticlesinteractingwithlipidmonolayersandbilayers,showingwithatimestepof2fsoncomputationalfacilitiessupportedbythetheeffectsofhydrophobicityandchargedensitiesofparticleNationalInstituteofSupercomputingandNetworking/KoreaInstitutesurfaces.Duanetal.’sall-atomsimulationsshowedthatcoronaofScienceandTechnologyInformationwithsupercomputingformationsuppressesinsertionandpenetrationofgrapheneresourcesincludingtechnicalsupport(KSC-2020-CRE-0095).oxidenanosheetsintolipidbilayers.40However,theeffectsofUmbrellaSamplingSimulations.Potentialsofmeanforce(PMFs)werecalculatedusingtheumbrellasamplingalgorithm,differentproteinsandmembranechargesonthebilayerwhichisacomputationalmethodtoovercomethepotentialbarrieranddynamicsandbindingstrengthhavenotyetbeensystematicallysampleallpossibleconfigurationsbyapplyingharmonicrestrainttothestudiedthroughall-atomsimulations.moleculeofinterestviaabiasedpotentialwithrespecttothereferenceAsafurthersteptowardunderstandingtheeffectsofvariousmolecule.61The10nm-sizedPSparticlecomplexedwithasingleplasmaproteinsandbilayerchargesonthebindingbetweenproteinortwoproteinswaspositionedabovethebilayerwithadistanceproteincoronaandbilayers,here,wereportall-atomMDof∼4nmbetweentheparticle−proteincomplexandbilayersurfacessimulationsofanionicPSparticlescomplexedwithasingleandthensolvatedwith∼170,000watermoleculesinaperiodicboxofsize16×16×29nm3.Thesystemswereneutralizedbyaddingplasmaproteinortwoplasmaproteins[humanserumalbumin(SA),IgG,andAPO]adsorbedontolipidbilayersasymmetri-counterionsandadditionalionsof0.15MNaClthatmimicsthephysiologicalcondition.Startingwithaninitialpositionoftheparticle−callycomposedofcytosolic(anionic)andextracellularproteincomplexwithadistanceof4nmfromthebilayersurface,the(zwitterionic)leaflets.Thebindingfreeenergiesarecalculatedparticle−proteincomplexwaspulledtowardthebilayersurfacewithafromumbrellasamplingsimulationsoftheparticlecomplexedforceconstantof1000kJmol−1nm−2,whichyieldsatotalof40samplewithproteinsmigratingtowardthezwitterionicoranionicconfigurations(calledwindows)withawindowspacingof0.1nmintheleaflet,showingtheeffectsofdifferentproteinsandbilayerdistanceof4nm(Figure1).These40windowswereequilibratedforchargesonthebindingstrengthbetweentheparticle−protein0.1nsandthenusedasstartingconfigurationsforumbrellasampling3752https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

2Langmuirpubs.acs.org/LangmuirArticlesystemsarelistedinTable1.Thelast10nstrajectorieswereusedtounbiasumbrellasamplingusingtheweightedhistogramanalysis62method.Errorswereestimatedfromthebootstrappinganalysis,calledtheBayesianbootstrappingofcompletehistograms,where63randomweightsareassignedtoallhistogramswithineachbootstrap.UnrestrainedSimulations.TheequilibratedfinalconfigurationsfromwindowshavingthelowestPMFvaluesintheabovementionedumbrellasamplingsimulationswereusedasstartingconfigurationsforunrestrainedsimulationsoftheparticle−proteincomplexinteractingwithlipidbilayers.Toobtainmoresamples,threesimulationsforeachsystemwerecarriedoutfor200ns(Table1),andthelast50nstrajectorieswereaveragedforanalyses.■RESULTSANDDISCUSSIONFree-EnergyCalculations:EffectsofDifferentProteinsandBilayerElectrostatics.ThestrengthsofthenonspecificbindingbetweenanionicPSparticle−proteincomplexesandlipidbilayers(asymmetricinnerandouterleaflets)werecomparedbycalculatingthebindingfreeenergies,aslistedinTable1.PMFsoftheparticle−proteincomplexmigratingtowardthebilayersurfacewereobtainedfromumbrellasamplingsimulationsof40windowsforeachsystem,whichwerecalculatedasafunctionofthedistancebetweencentersofmass(COMs)oftheparticle−proteincomplexandbilayerinFigure1.Potentialsofmeanforce(PMFs)forapolystyrene(PS)thebilayernormaldirectionandthenusedtocalculatefreeparticle(1stcolumn)oraPSparticle−proteincomplex(2nd∼4thcolumns)migratingtoward(a)anextracellular(zwitterionic)or(b)aenergiesofbinding(Figure1).Tounderstandtheeffectofcytosolic(anionic)leafletofthelipidbilayerasafunctionofthebilayercharges,bothcytosolic(anionic)andextracellulardistancebetweenparticle−proteincomplexandbilayercentersinthe(zwitterionic)leafletsofthelipidbilayerwereconsideredforbilayernormaldirection.Snapshotsforwindowshavingthelowestfree-energycalculations.InFigure1a,thelowestPMFsarePMFvaluesarepresentedforallsystems,whileinitialsnapshotsarefoundnearthebilayersurfaceformostsimulations,butthoseshownforsomesystems.Proteins,PSparticles,andtheiranionicPMFvaluesdiffer.Foraparticleorparticle−proteincomplexterminiare,respectively,representedasgreenribbonsandgrayandredmigratingtowardthezwitterionicleaflet,freeenergies(ΔG)aredots,whilebilayersandtheirheadgroupsarecoloredinlightanddarkmuchlowerforaparticlewithoutproteins(−25.5±1.0kJ/mol)blue,respectively.TheimageswerecreatedusingVisualMolecular67thanforparticlescomplexedwithSA,IgG,andAPODynamics.(respectively,−6.3±1.2,−2.5±3.9,and−14.5±1.6kJ/mol),showingastrongerinteractionofthebilayerwithbaresimulations.Eachwindowwassimulatedfor20ns,leadingtoatotalofparticlesurfacesthanwithproteinsontheparticlesurface.This800nsfor40windowsofeachparticle−proteinsystem.SimulatedindicatesthatproteincoronamoreweaklybindstothebilayerTable1.ListofSimulationsnumberofproteinsno.ofsimulationssimulationbilayersurfaceSAIgGAPO(orwindowsforPMF)time(ns)umbrellaZwitterionicleaflet40800sampling140800(PMF)1408001408001140800anionicleaflet408001408001408001408001140800unrestrainedZwitterionicleaflet3200simulation132001320013200113200anionicleaflet32001320013200132001132003753https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

3Langmuirpubs.acs.org/LangmuirArticlethandobareparticlesandthuscansuppressthebindingstrengthofthebindingofSAandIgGtothezwitterionicleafletbetweenPSparticlesandbilayersurfaces,inagreementwithmayberelativelyweak.Tocheckthis,windowshavingthe11,23,24experimentswithmodelmembranes.Inparticular,freelowestPMFswerefurtherequilibratedwithoutrestraintsfor200energiesarelowerforAPOthanforSAandIgG,indicatingans(Table1).ThreeunrestrainedsimulationswereperformedstrongerinteractionofthebilayerwithAPOthanwithSAandforeachsystemsothatmoresamplesareobtained.Figure3IgG,whichimpliestheeffectofdifferentproteinsontheproteinshowsthedistancebetweenPSparticle(orproteinforthecorona−bilayerbinding.particle−proteincomplex)andbilayercentersinthebilayerForaparticleorparticle−proteincomplexmigratingtowardnormaldirection.Particleswithoutproteinsbindtotheananionicleaflet,aparticlewithoutproteinsdoesnotshowazwitterionicleafletforwholesimulationtime,whiletheydonegativefree-energyvalue(Figure1b),indicatingthatannotinteractwiththeanionicleafletandsubstantiallymigrateanionicPSparticledoesnotbindtotheanionicleaflettowardbulkwater,whichoccursfordifferentorientationsofapparentlybecauseofelectrostaticrepulsion,asexpected.Freeparticles.Fortheparticle−proteincomplex,distancesarelowerenergiesaremuchlowerforparticlescomplexedwithSA,IgG,foranionicleafletsthanforzwitterionicleaflets,showingandAPO(respectively,−14.4±2.7,−12.3±0.7,and−34.3±strongerinteractionsofproteinswithanionicleafletsthanwith2.3kJ/mol)thanforabareparticle(0kJ/mol),indicatingthatzwitterionicleaflets,consistentwithfree-energycalculationsinproteincoronacanpromotethebindingbetweenparticlesandFigure1.Inparticular,SAandIgGinitiallyboundtotheanionicanionicbilayers.Inparticular,freeenergiesarelowerfortheleafletstayonthebilayersurfaceforwholesimulationtime,particle−proteincomplexesboundtoanionicleafletsthanforwhilethoseinitiallyboundtothezwitterionicleafletsometimesthoseboundtozwitterionicleaflets,implyingastrongermigratetowardthebulk-waterregion,indicatingtherelativelyinteractionofproteincoronawiththeanionicleafletthanwithweakbindingbetweenthoseproteinsandzwitterionicleaflets,thezwitterionicleaflet.Thisdependenceonthebilayerconsistentwithfree-energycalculations.TheseindicatethatelectrostaticsagreeswellwithexperimentsshowingtheAPOproteinsstronglybindtobothanionicandzwitterionicadsorptionofproteinsontotheanionicbilayerbutnotontoleaflets,whileSAandIgGstronglybindtoonlytheanionic64thezwitterionicbilayer.leaflet,againconfirmingtheeffectsofdifferentproteinsandUnrestrainedSimulations:InteractionsbetweenPar-bilayerchargesonthenonspecificbindingbetweentheparticle−ticle−ProteinComplexesandBilayers.Toquantifyproteincomplexandlipidbilayer.particle,protein,andbilayerconfigurations,massdensitiesTounderstandtheinteractionsbetweenparticle−proteinwerecalculatedforwindowshavingthelowestPMFvalues.complexes(orbareparticles)andbilayers,thenumberofFigure2showsthatanionicPSparticlesbindtothezwitterionichydrogenbondsbetweenproteins(orparticles)andlipidleafletbutdonotinteractwiththeanionicleaflet.Fortheheadgroupswascalculatedasafunctionoftime.Here,weparticle−proteincomplex,proteinsbindtothebilayersurfaceassumethatahydrogen-bondinginteractionexistswhentheandthuspreventPSparticlesfromdirectlybindingtothedonor−acceptordistanceis<0.35nmandtheangleofthe65bilayer.NotethatfreeenergiesarehigherforSAandIgGboundhydrogen-donor−acceptortripletis<30°.Notethatothertothezwitterionicleafletthanforthoseboundtotheanioniccriteriawiththedistancefrom0.3to0.4nmandtheanglefromleafletandforAPOboundtoanyleaflets,indicatingthatthe20to40°producesimilarqualitativetrends,confirmingthattheanalysisdoesnotsignificantlydependonthedistanceandanglecriteria.PSparticleswithoutproteinsformveryfewhydrogenbondswithbilayersurfaces,whileproteinsontheparticleformmanymorehydrogenbondswiththeanionicleafletthanwiththezwitterionicleaflet(Figure4),indicatingstrongerhydrogen-bondinteractionsofproteinswiththeanionicleafletthanwiththezwitterionicleaflet.Tofurtherunderstandtherolesofindividualresiduesofproteins,thenumberofhydrogenbondsbetweenlipidheadgroupsandproteinresiduessuchascationic(ArgandLys),anionic(AspandGlu),andotherresidueswascalculated.Figure5showsthatlipidheadgroupsformmorehydrogenbondswithcationicresiduesofproteinsthanwithanionicresiduesofproteins,implyingthattheprotein−bilayerbindingismodulatedbycationicresiduesofproteinsratherthanbyanionicresiduesofproteins.Anionicresiduesofproteinsformhydrogenbondswithanionicleafletsbutnotwithzwitterionicleafletspresumablybecauseproteinsmoretightlyinteractwithanionicleafletsthanwithzwitterionicleaflets,whichallowsanionicresiduestobeattractedtoanionicleafletsandformhydrogenbondswithlipidheadgroups.Thesechargeinteractionswerefurtherconfirmedbycalculatingradialdistributionfunctions(RDFs)betweenchargedaminoacidsandlipidheadgroups.InFigure6,forthezwitterionicleaflet,therearesharpRDFpeaksfortheinteractionsbetweencationicproteinresiduesandanioniclipidheadgroupsbutnotfortheinteractionsbetweenanionicproteinresiduesandcationiclipidFigure2.MassdensityprofilesofPSparticles,proteins,lipidheadgroups.RDFsofcationicproteinresiduesarehigherfortheheadgroups,andwater.anionicleafletthanforthezwitterionicleaflet.Theseresults3754https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

4Langmuirpubs.acs.org/LangmuirArticleFigure3.DistancesbetweenPSparticleandbilayercenters(1stcolumn;PSparticleswithoutproteins)andthosebetweenproteinandbilayercenters(2nd∼4thcolumns;PSparticleswithproteins)inthebilayernormaldirection,asafunctionoftime.Sincethreesimulationswereperformedforeachsystem,threedifferentcolorsareshown.Figure5.Numberofhydrogenbondsbetweenlipidheadgroupsandaminoacidsofproteinssuchascationicresidues(ArgandLys),anionicresidues(AspandGlu),andotherresidues.Figure4.Numberofhydrogenbondsbetweenproteinsandlipid(Table1andFigure7).Freeenergiesare−11.6±3.2and−24.8headgroupsofzwitterionic(left)andanionic(right)leafletsasa±0.8(×10−8)cm2s−1,respectively,forzwitterionicandanionicfunctionoftime.Sincethreesimulationswereperformedforeachleaflets,showingstrongerinteractionsofproteinswiththesystem,threedifferentcolorsareshown.anionicleafletthanwiththezwitterionicleaflet.RecallfromFigure1thatfortheparticlecomplexedwithasingleSAorIgG,indicatethattheproteincorona−bilayerbindingismodulatedfreeenergiesare−6.3to−2.5and−14.4to−12.3(×10−8)cm2bychargeinteractionsbetweencationicresiduesofproteinsands−1forzwitterionicandanionicleaflets,respectively.Theseshowanionicheadgroupsoflipids,whichisstrongerfortheanioniclowerfreeenergiesfortheparticlecomplexedwithtwoproteinsleafletthanforthezwitterionicleaflet.thanfortheparticlecomplexedwithasingleproteinapparentlyEffectofProteinDensityontheProteinCorona−becauseofthehigherextentoftheprotein−bilayerinteractionBilayerBinding.Tounderstandtheeffectofproteindensityonformedbytwoproteinsthanbyasingleprotein.Tomorethebindingstrengthbetweentheparticle−proteincomplexandpreciselycomparetheirbindingstrengths,windowshavingthethebilayersurface,umbrellasamplingsimulationswerelowestPMFswerefurtherequilibratedwithoutrestraintsfor200performedfortheparticlecomplexedwithtwoproteins(SAns.Toobtainmoresamples,threesimulationswereperformedandIgG)migratingtowardthezwitterionicoranionicleafletforeachsystem.Figure8showsthatdistancesbetweenproteins3755https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

5Langmuirpubs.acs.org/LangmuirArticleFigure8.DistancebetweeneachproteinandbilayercentersintheFigure6.Radialdistributionfunctionsbetweenanionicresidues(Aspbilayernormaldirection,asafunctionoftime.SincethreesimulationsandGlu)ofproteinsandcationicheadgroups(cholineandamine)ofwereperformedforeachsystem,threedifferentcolorsareshown.lipids(toprow)andbetweencationicresidues(ArgandLys)ofproteinsandanionicheadgroups(phosphateandcarboxylate)oflipids(bottomrow).Figure9.NumberofhydrogenbondsbetweenproteinsandlipidFigure7.PMFsforaPSparticlecomplexedwithSAandIgGmigratingheadgroupsofzwitterionic(toprow)andanionicleaflets(bottomrow)towardazwitterionic(left)oranionicleaflet(right)ofthelipidbilayerasafunctionoftime.Sincethreesimulationswereperformedforeachasafunctionofthedistancebetweentheparticle−proteincomplexandsystem,threedifferentcolorsareshown.bilayercentersinthebilayernormaldirection.SnapshotsforwindowshavingthelowestPMFvaluesarepresented.boundtothezwitterionicoranionicleafletwerecalculatedusing66andanionicleafletsareverylowanddonotfluctuatemuchfortheDSSPprogram.Figure10showsthatproteinsboundtowholesimulationtime,whereasoneofthesimulationswiththezwitterionicleafletretaintheirhelicalstructure(100thtozwitterionicleafletsshowssubstantialmigrationoftheparticle−130thresidues),whilethoseboundtotheanionicleafletshowproteincomplextowardbulkwater,similartoresultsfromthereducedhelicitypresumablybecauseproteinscomplexedsimulationsoftheparticlecomplexedwithasingleprotein.Thewiththeparticleinteractmorestronglywiththeanionicleafletnumberofhydrogenbondsbetweenproteinsandlipidthanwiththezwitterionicleaflet,whichdisruptsthehelicalheadgroupswasalsocalculated(Figure9),whichshowsthatstructureofproteinsboundtotheanionicleaflet.ThisindicatesSAandIgGformmorehydrogenbondswithanionicleafletsthattheproteinstructurecanbeinfluencedbystrongchargethanwithzwitterionicleaflets,againconfirmingstrongerinteractionsbetweenproteinsandbilayers,toanextentinteractionsofproteinswithanionicleafletsthanwithdependentonbilayercharges.zwitterionicleaflets.Inparticular,thenumbersofhydrogenExperimentally,Montisetal.observedthatthelateralbondsforasingleprotein(Figure4)andtwoproteins(Figure9)dynamicsofmodelmembranesdecreaseswheninteracting22donotsignificantlydiffer,showinglittleeffectofproteindensitywithproteincorona,althoughanaccurateaccountingofthisonthebindingbetweenproteinsandlipidbilayers.phenomenonhasnotyetbeenunderstoodattheatomicscale.EffectofProteinCoronaontheProteinStructureandToresolvethis,lateraldiffusioncoefficientsoflipidswereBilayerDynamics.Asdiscussedabove,theparticle−proteincalculatedfromtheslopesofthemean-squaredisplacementsofcomplexesbindtolipidbilayersviahydrogen-bondinteractionslipid-phosphateatomsinthexy-plane(thedirectionperpen-betweenchargedgroupsofproteinsandbilayerheadgroups,diculartothebilayernormal).Forcomparison,wefirstwhichmayinfluencetheproteinstructureandbilayerdynamics.calculatedlateraldiffusioncoefficientsofthebilayerboundtoTounderstandtheeffectoftheproteincorona-bilayerbindingPSparticleswithoutproteins,givingthevaluesof1.7to2.2ontheproteinstructure,secondarystructuresofAPOproteins(×10−8)cm2s−1forthezwitterionicleafletand2.1to2.33756https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

6Langmuirpubs.acs.org/LangmuirArticleFigure10.SecondarystructureprofilesofAPOproteins(100thto150thresidues)boundtozwitterionicleaflets(left)andanionicleaflets(right)asafunctionoftime.Sincethreesimulationswereperformedforeachsystem,threestructureprofilesarepresented.(×10−8)cm2s−1fortheanionicleaflet(Table2).NotethatPSthatanionicPSparticlesdonotdisruptthelipidbilayer.23Inparticlesdonotbindtotheanionicleaflet,andhence,theparticular,Table2showsthatlateraldiffusivitiesofthezwitterionicleafletdecreasewhenboundtoproteinspresumablyabecausehydrogenbondsbetweenproteinsandlipidheadgroupsTable2.LateralDiffusionCoefficients(DL)ofLipidsrestrictthemotionoflipids,inagreementwithMontisetal.’sbindingtothebilayerD(10−8cm2s−1)Lexperimentsshowingthattheproteincorona−bilayerbindingZwitterionicnoprotein√1.7±0.1inducestheraft-like(ordered)phaseofthebilayer,leadingtoaleaflet√2.2±0.2decreaseinthelateraldynamics.22Incontrast,lateraldiffusivities√2.0±0.3oftheanionicleafletincreasewhenboundtoproteinsSA√1.3±0.1presumablybecauseproteinsinteractmoretightlywithanionic√1.4±0.4leafletsthanwithzwitterionicleafletsandthussignificantly1.9±0.1disorderanionicleaflets.NotethatlargedifferencesbetweenIgG√0.9±0.2membranecomponents,proteinconcentrations,particlesizes,√1.0±0.3andmasstransportconditionsofthesimulationscomparedto2.0±0.3thoseinexperimentsprecludeanyquantitativecomparisonAPO√1.2±0.4betweenthetwo.Also,itwouldobviouslybeinterestingto√1.5±0.2investigatecooperativeeffectsofmultipleproteinsinsoftand√1.1±0.4hardcoronas,whichisbeyondthescopeofthispaperandhopeanionicnoprotein2.3±0.1toreportonelsewhere.Despitethis,wehaveclearlyshownthatleaflet2.1±0.4barePSparticlesformveryfewhydrogenbondswithlipid2.2±0.2headgroupsandthusdonotinfluencethelateraldynamicsofSA√2.9±0.1bilayers,whileproteinsontheparticlesurfaceformhydrogen√2.2±0.1bondswithlipidheadgroupsandthusinfluencethelateral√2.4±0.2dynamicsofbilayers.Inparticular,hydrogenbondsbetweenIgG√2.7±0.4proteinsandzwitterionicleafletsrestrictthemotionoflipidsand√2.7±0.1thusreducetheirlateraldynamics,whilethetightbinding√3.3±0.3betweenproteinsandanionicleafletsdisruptslipidsandthusAPO√2.5±0.1increasestheirlateraldynamics,showingthedependenceon√2.0±0.1bilayercharges.Thesefindingshelpexplaintheexperimental√2.9±0.3aobservationsregardingthefactthatthelateraldynamicsoflipidSincethreesimulationswereperformedforeachsystem,threevalues22bilayersdecreaseswheninteractingwithproteincoronaandarerepresented.alsosuggestthatthiseffectofproteincoronaonthebilayerdynamicsdependsonbilayercharges.diffusivityvaluesof2.1to2.3(×10−8)cm2s−1canbealsoconsideredforpurebilayerswithoutparticlesandproteins.This,■combinedwithhydrogen-bondinteractions,indicatesthatCONCLUSIONSalthoughanionicPSparticlesbindtothezwitterionicleaflet,AnionicPSparticlescomplexedwithasingleplasmaproteinortheyformveryfewhydrogenbondswithlipidheadgroupsandtwoplasmaproteins(SA,IgG,andAPO)weresimulatedwiththuscannotsignificantlyinfluencethelateraldynamicsofthethelipidbilayerasymmetricallycomposedofanionic(cytosolic)bilayer,asobservedinDiSilvioetal.’sexperimentsthatshowedandzwitterionic(extracellular)leaflets.Tocomparethebinding3757https://doi.org/10.1021/acs.langmuir.1c00249Langmuir2021,37,3751−3760

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