Brain UDPgalactose: ceramide galactosyltransferase Purification of a catalytically active protein obtained after proteolytic digestion.
A procedure for the purification of UDPgalactose--2-hydroxyacylsphingosine galactosyltransferase (EC 2.4.1.45) including detergent extraction, ion-excharge chromatography and proteolytic digestion was developed. The active fraction obtained by this procedure had about 100 times higher specific activity than microsomes. Enzymic activity resisted destruction by pronase treatment at 4 degrees C. Agarose gel chromatography indicated the presence of an enzyme-phospholipid-detergent complex with a molecular weight between 400 000 and 500 000. Intact phospholipids seemed to be required for full enzymic activity as evidenced by the drastic loss of activity upon treatment with phospholipase A or C.
D000818:Animals; D001921:Brain / Q000201:enzymology*; D004355:Drug Stability; D005700:Galactosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology; D010740:Phospholipases; D011402:Pronase
D005700:Galactosyltransferases; D010740:Phospholipases; D011402:Pronase