Effect of phospholipid composition on the surface potential of liposomes and the activity of enzymes incorporated.
1. Microsomes of rat liver and brain and mitochondria of rat liver and guinea-pig brown adipose tissue were solubilized with the nonionic detergent Lubrol-WX and the solubilized material was incorporated into liposomes of various phospholipid composition. In proteoliposomes thus formed the kinetics of arylsulphatase, glycerol-3-phosphate dehydrogenase, monoamine oxidase and acetylcholinesterase were measured. 2. It was shown that the apparent Km values of arylsulphatase and glycerol-3-phosphate dehydrogenase were higher in liposomes prepared with negatively charged phospholipids and lower in liposomes containing positively charged organic amines, as compared with th Km value of enzymes incorporated into liposomes prepared from phosphatidylcholine alone. The opposite was true for monoamine oxidase and acetylcholinesterase, i.e. enzymes possessing cationic substrates. Phospholipid composition did not essentially influence the activity of the enzymes extrapolated for infinite substrate concentration (V values). 3. As compared with proteoliposomes made from phosphatidylcholine, the binding constant (Ka) of 8-anilino-1-naphthalene sulphonate was higher when the vesicles contained acidic phospholipids or bis(hexadecanyl)phosphate and lower when they contained organic amines. 4. A correlation between changes of the surface potential calculated from Ka values of anilino-naphthalene sulphonate and variations in apparent Km values of the four enzymes under investigation indicates that the activity of membrane-bound enzymes may be modulated by charged phospholipids due to decreasing or increasing substrate concentration in the unstirred layer, as predicted from the Boltzmann distribution.
D000110:Acetylcholinesterase / Q000378:metabolism*; D002001:Adipose Tissue, Brown / Q000378:metabolism; D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism*; D001921:Brain / Q000378:metabolism; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism*; D006168:Guinea Pigs; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism*; D008564:Membrane Potentials; D008861:Microsomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism; D008995:Monoamine Oxidase / Q000378:metabolism*; D051381:Rats; D013429:Sulfatases / Q000378:metabolism*
D008081:Liposomes; D005993:Glycerolphosphate Dehydrogenase; D008995:Monoamine Oxidase; D000110:Acetylcholinesterase; D013429:Sulfatases; D001192:Arylsulfatases