UDPglucose-ceramide glucosyltransferase from porcine submaxillary glands is associated with the Golgi apparatus.
Subcellular distribution of pig submaxillary gland UDPglucose-ceramide glucosyltransferase (EC 2.4.1.80), the enzyme which catalyses the first step during the sequential addition of carbohydrate moieties for ganglioside biosynthesis, was studied. The results presented strongly suggest that in pig submaxillary gland, the transfer of glucose on endogenous or exogenous ceramides takes place in the Golgi apparatus: the specific activity of UDPglucose-ceramide glucosyltransferase increased in parallel with the activity of a known marker of the Golgi apparatus, UDPgalactose-ovomucoid galactosyltransferase. The specific activity of the glucosyltransferase was 18-times higher in the purified Golgi membranes than in the postnuclear supernatant and the yield was over 30%. An apparent Km of 22 microM for UDPglucose and 54 microM for ceramides was determined. Maximal glucosylation of endogenous ceramides was achieved at pH 6.5 in the presence of NADH (1 mM) as inhibitor of pyrophosphatases and with Mn2+ (5 mM). It was found that the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps) is an efficient activator for the glucosylation of exogenous ceramides.
Coste
H
H
Martel
M B
MB
Azzar
G
G
Got
R
R
D000818:Animals; D002855:Chromatography, Thin Layer; D005964:Glucosyltransferases / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D013363:Submandibular Gland / Q000648:ultrastructure*; D013552:Swine; D013997:Time Factors
D008345:Manganese; D005964:Glucosyltransferases; C023246:ceramide glucosyltransferase; D008274:Magnesium