Interaction of membranous enzymes with membranous lipid substrates. Hydrolysis of diacylglycerol by lipase in rat brain microsomes.
The phospholipids in rat brain microsomes were labeled with tritium by intracerebral administration of radioactive fatty acids and converted to diacylglycerol with phospholipase C. The latter lipid was hydrolyzed in situ at pH 4.8, to monoacylglycerol and fatty acid by the endogenous microsomal lipase. This paper provides an experimental approach to determine whether the lipid was degraded by enzyme molecules residing in its own membrane (intramembrane interaction) or an adjacent membrane (intermembrane interaction). Direct interaction between separate membranes containing enzyme or substrate showed the existence of the inter-membrane route while dilution experiments provided evidence for the presence of the intramembrane interaction as well. A probable difference in the mechanisms of these two interactions is suggested by different shapes of the curves that describe the reaction rate as a function of the endogenous substrate. The curve resulting from the intermembrane interaction was hyperbolic while that representing the intramembrane route was of a parabola-like shape. Competition experiments suggested that when given a choice between the two, the enzyme utilized preferentially the substrate molecules in its own membrane.
D000818:Animals; D001921:Brain / Q000201:enzymology*; D004075:Diglycerides; D006868:Hydrolysis; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008563:Membrane Lipids; D008861:Microsomes / Q000201:enzymology*; D010743:Phospholipids; D051381:Rats
D004075:Diglycerides; D008563:Membrane Lipids; D010743:Phospholipids; D008049:Lipase