title: Mitochondrial dolichyl-phosphate mannose synthase. Purification and immunogold localization by electron microscopy.
abstract: 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.
authors.affiliation: University of Lyon, Lyon-Sud Medical School, Department of Biochemistry, France.
authors.forename: F
authors.initials: F
authors.lastname: Gasnier
authors.forename: R
authors.initials: R
authors.lastname: Rousson
authors.forename: F
authors.initials: F
authors.lastname: Lerme
authors.forename: E
authors.initials: E
authors.lastname: Vaganay
authors.forename: P
authors.initials: P
authors.lastname: Louisot
authors.forename: O
authors.initials: O
authors.lastname: Gateau-Roesch
mesh_terms: 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
chemical_list: D007074:Immunoglobulin G; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase