title: Large-scale expression of recombinant sialyltransferases and comparison of their kinetic properties with native enzymes.
abstract: Values of Km were determined for three purified sialyltransferases and the corresponding recombinant enzymes. The enzymes were Gal beta 1-4GlcNAc alpha 2-6 sialyltransferase and Gal beta 1-3(4)GlcNAc alpha 2-3 sialyltransferase from rat liver; these enzymes are responsible for the attachment of sialic acid to N-linked oligosaccharide chains; and the Gal beta 1-3GalNAc alpha 2-3 sialyltransferase from porcine submaxillary gland that is responsible for the attachment of sialic acid to O-linked glycoproteins and glycolipids. A procedure for the large scale expression of active sialyltransferases from recombinant baculovirus-infected insect cells is described. For the liver enzymes values of Km were determined using rat and human asialo alpha 1 acid glycoprotein and N-acetyllactosamine as variable substrates; lacto-N-tetraose was also used with the Gal beta 1-3(4)GlcNAc alpha 2-3 sialyltransferases. Antifreeze glycoprotein was used as the macromolecular acceptor for the porcine enzyme. Values for Km were also determined using CMP-NeuAc as the variable substrate.
authors: Cytel Corporation, San Diego, California 92121, USA.
authors: M A
authors: MA
authors: Williams
authors: H
authors: H
authors: Kitagawa
authors: A K
authors: AK
authors: Datta
authors: J C
authors: JC
authors: Paulson
authors: J C
authors: JC
authors: Jamieson
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005822:Genetic Vectors; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012799:Sialyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D012799:Sialyltransferases