membrane rafts detected by a peptidyl molecular probe - Matsubara et al. - Unknown - Unknown

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SupportingInformationHeterogeneousganglioside-enrichednanoclusterswithdifferentdensitiesinmembraneraftsdetectedbyapeptidylmolecularprobeTeruhikoMatsubara*,KazutoshiIIjima,TakahiroKojima,MiwaNakamura,ErikaMiyamoto,andToshinoriSato*-1-

1TableS1.Occupiedareas(%)ofdomainsintheGM1/SM/cholesterol/POPC(X:10:35:50)membrane(n=3)areaordomainX=0X=2X=5X=10areaa64.6±0.162.1±5.056.1±1.846.8±0.9domainb35.4±0.137.9±5.034.5±5.035.4±3.2domaing(notdetected)(notdetected)9.4±3.617.9±2.3TableS2.Averagediameters(nm)ofdomaingintheGM1/SM/cholesterol/POPC(X:10:35:50)membraneX=0X=2X=5X=10average±standarddeviations(notdetected)62.7±0.467.2±6.062.7±1.1numberofdomains(n)–163651TableS3.AffinitiesofCTBandtheGCBP–avidincomplexfortheGM1monolayerassessedbytheABCmethodCP8–avidincomplexCTBGCBP–avidincomplex(control)Kd2.3±0.4151±11128±38DAmax0.30±0.174.3±1.21.9±1.0FigureS1.Typicalplotsof[protein]versus[protein]/absorbanceforassessmentsofKdvalues.-2-

2TableS4.MolecularspeciesofGM1fromthebovinebraindetectedbyLC-MS.m/zm/zretentionarearelativeareagangliosideiontypeceramidetype(calcd.)(found)time(min)(´106)(%)GM1[M–H]–1544.91545.5d18:1–C18:030.61.7352.0GM1[M–H]–1572.91573.6d20:1–C18:030.61.6048.0TableS5.TypicalacidicmolecularspeciesfromPC12cellsdetectedbyLC-MS.ganglio-m/zm/zretentionarearelativeiontypeceramidetypeside(calcd.)(found)time(min)(´106)area(%)GM1[M–H]–1516.81516.9d16:1–C18:0(d18:1–C16:0)30.30.2718.3[M–H]–1544.91544.8d18:1–C18:030.10.1711.2[M–H]–1572.9–d20:1–C18:0–N.D.*0.0[M–H]–1600.91599.1d22:1–C18:0(d20:1–C20:0)29.01.0670.6GD1a[M–2H]2–903.5903.8d16:1–C18:0(d18:1–C16:0)32.04.6544.1[M–2H]2–917.5917.7d18:1–C18:031.90.757.1[M–2H]2–931.5931.5d20:1–C18:031.91.0610.1[M–2H]2–945.5945.3d22:1–C18:0(d20:1–C20:0)31.71.2011.4[M–2H]2–959.5959.5d24:1–C18:031.71.4013.3[M–2H]2–958.5958.7d24:1–C18:131.71.4814.1GT1b[M–2H]2–1049.01049.5d16:1–C18:0(d18:1–C16:0)32.61.6321.9[M–2H]2–1063.01063.7d18:1–C18:032.50.344.5[M–2H]2–1077.01077.0d20:1–C18:032.70.689.2[M–2H]2–1091.01091.1d22:1–C18:0(d20:1–C20:0)32.60.739.8[M–2H]2–1105.11105.1d24:1–C18:032.63.8551.5[M–2H]2–1104.11105.1d24:1–C18:132.50.243.2GQ1b[M–2H]2–1194.61194.5d16:1–C18:0(d18:1–C16:0)32.60.4422.9[M–2H]2–1208.6–d18:1–C18:0–N.D.0.0[M–2H]2–1222.61223.0d20:1–C18:032.70.189.3[M–2H]2–1236.61236.5d22:1–C18:0(d20:1–C20:0)32.60.2814.3[M–2H]2–1250.61250.9d24:1–C18:032.71.0353.5*N.D.,notdetected(area,<1´104).-3-