A Monolayer Assay Tailored to Investigate Lipid–Protein Systems

A Monolayer Assay Tailored to Investigate Lipid–Protein Systems

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时间:2019-08-06

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1、CHEMPHYSCHEMARTICLESDOI:10.1002/cphc.201300035AMonolayerAssayTailoredtoInvestigateLipid–ProteinSystems[b][a]GrzegorzChwastekandPetraSchwille*Modelmembranesystemshavebecomeinvaluabletoolstoin-miniaturisedmonolayerassaythatcaneasilybecombinedvestigatespecificfeaturesofcellularmembranes.Althoughwithst

2、andardanalyticaltechniquessuchasconfocalfluores-avarietyofdifferentexperimentalassaysdoesexist,manyofcencemicroscopyandfluorescencecorrelationspectroscopythemarerathercomplicatedintheirpreparationanddifficult(FCS).intheirpracticalrealisation.Here,weproposeanewsimple1.IntroductionForthelast30years,b

3、iologicalmembraneshavebeenkeyob-layer.Thissystemlacksessentialfeaturesofabiologicalmem-jectsofbiophysicalresearch.Besidestheobviousinvivostud-brane,asitconsistsofonlyonelayeroflipidsdepositedatiesonnativecellularmembranes,alargenumberofinvitroawater–airinterface.However,itprovidesquantitativeaccess

4、methodsandassayshasbeendeveloped,inordertoquantita-toparameterswhichcanhardlybetunedinbilayerassays,buttivelyinvestigatebiologicalprocessesunderdefinedandwell-whichareneverthelessofgreatrelevanceforeverypotentialcontrolledconditions.Themostrecentamongtheseap-interactionofproteinswiththemembrane,suc

5、hasthepack-proachescanbesummarisedunderthenotationofso-calledingdensityoflipids.Whilemonolayershavebeenvalidatedfor[1][10]“minimalbiologicalsystems”thataresupposedtoretaintheprotein-membranebindingassays,theirapplicationsarestillcoreofabiologicalphenomenoninspiteofthedramaticallylimitedbyseveralins

6、trumentalfeatures.Mostimportantly,con-reducedcompositionalandtopologicalcomplexity.Somepow-ventionalLangmuirtroughs,unlessmodified,arenotcompati-erfulandwidelyappliedminimalmembranesystemsare,forblewithconfocalmicroscopes,restrictingtheirusemostlyto[2–4]example,giantunilamellarvesicles(GUVs),blackl

7、ipidmem-themonitoringofthesurfacepressure(SP).Additionally,[5][6,7]branes,supportedlipidbilayers(SLBs)andlipidmonolay-asinglemonolayerexperimentrequiresusuallymorethan[10–13]ers.Usingthesesystems,interactio

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