Partial purification and properties of a murine plasmacytoma glucosyltransferase.
The enzyme UDP-glucose:dolichylphosphate glucosyltransferase has been purified 1700-fold from MOPC-315 plasmacytoma tissue. The purification combines differential detergent extraction of purified rough endoplasmic reticulum with subsequent ion exchange chromatography, dye affinity chromatography, and hydroxylapatite chromatography of the extract. The partially purified glucosyltransferase exhibits a Km of 0.79 microM for UDP-Glc and a Km of 0.65 microM for dolicholphosphate in the presence of 4 mg/ml of phosphatidylcholine. The reaction is dependent upon the addition of exogenous dolicholphosphate. The enzyme is activated by the choline containing lipids phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin. The dye Remazol blue acts as a competitive inhibitor of the enzyme with respect to UDP-Glc. The molecular weight of the enzyme has been determined to be approximately 37,000. The sole reaction product has been identified as dolichylphosphate glucose by isolation of the product by DEAE-cellulose chromatography and subsequent analysis of the acid-hydrolyzed product by both Bio-Gel P2 gel filtration and paper chromatography.
D000818:Animals; D002460:Cell Line; D004492:Edetic Acid / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000201:enzymology; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D010743:Phospholipids / Q000494:pharmacology; D010954:Plasmacytoma / Q000201:enzymology*; D012995:Solubility
D010743:Phospholipids; D004533:Egtazic Acid; D004492:Edetic Acid; D005964:Glucosyltransferases; C033917:UDPglucose dolicholphosphate glucosyltransferase