Galactose oxidase action on galactose containing glycolipids - a fluorescence method

M Fortelius, Peter Mattjus

    Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

    5 Citeringar (Scopus)

    Sammanfattning

    Features that alter the glycolipid sugar headgroup accessibility at the membrane interface have been studied in bilayer lipid model vesicles using a fluorescence technique with the enzyme galactose oxidase. The effects on oxidation caused by variation in the hydrophobic moiety of galactosylceramide or the membrane environment for galactosylceramide, monogalactosyldiacylglycerol and digalactosyldiacylglycerol were studied. For this study we combined the galactose oxidase method for determining the oxidizability of galactose containing glycolipids, and the fluorescence method for determining enzymatic hydrogen peroxide production. Exposed galactose residues with a free hydroxymethyl group at position 6 in the headgroup of glycolipids were oxidized with galactose oxidase and subsequently the resultant hydrogen peroxide was determined by a combination of horseradish peroxidase and 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red). Amplex Red reacts with hydrogen peroxide in the presence of horseradish peroxidase with a 1:1 stoichiometry to form resorufin. With this coupled enzyme approach it is also possible to determine the galactolipid transbilayer membrane distribution (inside-outside) in bilayer vesicles.
    OriginalspråkOdefinierat/okänt
    Sidor (från-till)103–110
    Antal sidor8
    TidskriftChemistry and Physics of Lipids
    Volym142
    Utgåva1-2
    DOI
    StatusPublicerad - 2006
    MoE-publikationstypA1 Tidskriftsartikel-refererad

    Nyckelord

    • Amplex Red
    • DGDG
    • enzyme
    • galactosylceramide
    • membrane
    • MGDG
    • phosphatidylcholine
    • sphingomyelin
    • vesicle

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