Galactosyltransferase activities in mitochondria outer membrane: biosynthesis of galactosylated proteins.
1. Mitochondria outer membranes prepared from mouse livers were purified on a discontinuous sucrose gradient. Control in electron microscopy and marker enzymes assays confirmed purity and homogeneity of this fraction. 2. Purified mitochondria outer membranes exhibited significant UDP-galactose: glycoprotein galactosyltransferase activities when incubated with endogenous or exogenous glycoprotein acceptors in presence of detergent (Nonidet P40). 3. Some properties of two distinct mitochondrial galactosyltransferases, acting respectively on ovomucoid and ovine asialo-mucin were investigated. 4. Transfer of galactose on ovomucoid was maximal for a pH of 7.6 at 33 degrees C whereas asialo-mucin galactosyltransferase exhibited an optimum pH of 5.6 for an optimal temperature of 46 degrees C. 5. These two distinct membrane-bound enzymes were both inhibited by diacylglycerophospholipids whereas lysophospholipids modulated both enzymes in a different way: at 5 mM lysophosphatidylcholine, asialo-mucin galactosyltransferase was slightly stimulated while ovomucoid galactosyltransferase was markedly activated. 6. The most important activating effect on ovomucoid galactosyltransferase was obtained with a phospholipid containing a long aliphatic side chain linked by an ester bond in sn-1 of glycerol, an hydroxyl group or hydrogen atoms in sn-2 and a phosphorylcholine head group in sn-3.
Gasnier
F
F
Department of Biochemistry, Lyon-Sud Medical School, University of Lyon, INSERM-CNRS U.189, Oullins, France.
Louisot
P
P
Gateau
O
O
D000818:Animals; D005700:Galactosyltransferases / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000201:enzymology* / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure
D006023:Glycoproteins; D005700:Galactosyltransferases