Mitochondrial dolichyl-phosphate mannose synthase. Purification and immunogold localization by electron microscopy.
Mitochondrial dolichyl-phosphate mannose synthase has been purified to homogeneity using an original procedure, reconstitution into specific phospholipid vesicles and sedimentation on a sucrose gradient as final step. The enzyme has an apparent molecular mass of 30 kDa on an SDS/polyacrylamide gel. Increased enzyme activity could be correlated with this polypeptide band. A specific antibody was raised in rabbits against this transferase. Specific IgG obtained from the immune serum removed enzymatic activity from a detergent extract of mitochondrial outer membrane and reacted specifically with the 30-kDa band on immunoblots. Furthermore, an immunocytochemical experiment proved the localization of dolichyl-phosphate mannose synthase on the cytosolic face of the outer membrane of mitochondria.
Gasnier
F
F
University of Lyon, Lyon-Sud Medical School, Department of Biochemistry, France.
Rousson
R
R
Lerme
F
F
Vaganay
E
E
Louisot
P
P
Gateau-Roesch
O
O
D000818:Animals; D015153:Blotting, Western; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D007074:Immunoglobulin G; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000201:enzymology; D008364:Mannosyltransferases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D051379:Mice; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight
D007074:Immunoglobulin G; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase