Lactose synthetase. The isolation and characterisation of the protein-protein complex.
Galactosyl transferase and alpha-lactalbumin were concurrently isolated from bovine milk. The galactosyl transferase had an S20,w of 3.0 S and D20,w of 60 mum2/s. It exists as a monomer of 46000 molecular weight, as determined by sedimentation equilibrium and dodecylsulphate-polyacrylamide electrophoresis. Aggregation of the enzyme was promoted by N-acetylglucosamine. Sedimentation velocity experiments show an association between galactosyl transferase and alpha-lactalbumin in the presence of N-acetylglucosamine. In contrast, UDP-galactose, UDP or lactose do not promote protein-protein association. A complex between galactosyl transferase and alpha-lactalbumin was isolated by gel filtration in the presence of excess alpha-lactalbumin and either N-acetylglucosamine or glucose. The complex was stable over a range of concentrations of these components. The complex is a discrete homogeneous entity with a molecular weight of 60000, corresponding to one molecule of galactosyl transferase and one molecule of alpha-lactalbumin. The estimated association constants for the ternary complexes of the two proteins and either of the sugars, suggest that alpha-lactalbumin enhances equally the binding of N-acetylglucosamine or glucose to the galactosyl transferase.
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D005260:Female; D005700:Galactosyltransferases / Q000302:isolation & purification; D007768:Lactalbumin / Q000302:isolation & purification; D007788:Lactose Synthase / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008892:Milk / Q000032:analysis / Q000201:enzymology; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation
D046911:Macromolecular Substances; D007768:Lactalbumin; D005700:Galactosyltransferases; D007788:Lactose Synthase