Dynamics of Ceramide Channels Detected Using a Microfluidic System 英文参考文献.docVIP

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Dynamics of Ceramide Channels Detected Using a Microfluidic System 英文参考文献.doc

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Dynamics of Ceramide Channels Detected Using a Microfluidic System 英文参考文献

DynamicsofCeramideChannelsDetectedUsinga MicrofluidicSystem ChenrenShao2.,BingSun1.¤,DonL.DeVoe2,MarcoColombini1* 1Department of Biology, University of Maryland, College Park, Maryland, United States of America, 2Department of Mechanical Engineering, University of Maryland, CollegePark,Maryland,UnitedStatesofAmerica Abstract Ceramide,aproapoptoticsphingolipid,hasbeenshowntoformchannels,inmitochondrialoutermembranes,largeenough to translocate proteins. In phospholipid membranes, electrophysiological studies and electron microscopic visualization bothreport thatthesechannelsforminarangeofsizeswithamodalvalueof10nmindiameter. Ahydrogen bonded barrel-likestructureconsistingofhundredsofceramidemoleculeshasbeenproposedforthestructureofthechanneland this is supported by electrophysiological studies and molecular dynamic simulations. To our knowledge, the mechanical strengthanddeformabilityofsuchalargediameterbutextremelythincylindricalstructurehasneverbeenreported.Here 3+ we present evidence for a reversible mechanical distortion of the cylinder following the addition of La . A microfluidic systemwasusedtorepeatedlylowerandthenrestoretheconductancebyalternativelyperfusingLa3+andEDTA.Although aspectsofthekineticsofconductancedropandrecoveryareconsistentwithadisassembly/diffusion/reassembly model, others are inconsistent with the expected time scale of lateral diffusion of disassembled channel fragments in the membrane. The presence of a residual conductance following La3+ treatment and the relationship between the residual conductanceandtheinitialconductancewerebothindicativeofadistortion/recoveryprocessinanalogywithapressure- induceddistortionofaflexiblecylinder. Citation: Shao C, Sun B, DeVoe DL, Colombini M (2012) Dynamics of Ceramide Channels Detected Using a Microfluidic System. PLoS ONE 7(9): e43513. doi:10.1371/journal.pone.0043513 Editor:MarekCebecauer,J.HeyrovskyInstituteofPhysicalChemistry,CzechRepublic ReceivedMarch28,2012;AcceptedJuly25,2012;P