Galactosylation of N-linked oligosaccharides by human beta-1,4-galactosyltransferases I, II, III, IV, V, and VI expressed in Sf-9 cells

Glycobiology. 2001 Oct;11(10):813-20. doi: 10.1093/glycob/11.10.813.


Several studies showed that Sf-9 cells can synthesize the galactosylated N-linked oligosaccharides if beta-1,4-galactosyltransferase (beta-1,4-GalT) is supplied. The full-length human beta-1,4-GalT I, II, III, IV, V, and VI cDNAs were independently transfected into Sf-9 cells, and the galactosylation of endogenous membrane glycoproteins was examined by lectin blot analysis using Ricinus communis agglutinin-I (RCA-I), which preferentially interacts with oligosaccharides terminated with Galbeta1-->4GlcNAc group. Several RCA-I-reactive bands appeared in all of the gene-transfected cells, and disappeared on treatment of blots with beta-1,4-galactosidase or N-glycanase prior to incubation with lectin. Introduction of the antisense beta-1,4-GalT II and V cDNAs separately into human colorectal adenocarcinoma SW480 cells, in which beta-1,4-GalT I, II, and V genes were expressed, resulted in the reduction of RCA-I binding toward N-linked oligosaccharides of the membrane glycoproteins. Differences were found in their K(m) values toward UDP-Gal and GlcNAcbeta-S-pNP and in their acceptor specificities toward oligosaccharides with the GlcNAcbeta1-->4(GlcNAcbeta1-->2)Man branch and with the GlcNAcbeta1-->6(GlcNAcbeta1-->2)Man branch. These results indicate that beta-1,4-GalTs II, III, IV, V, and VI are involved in the N-linked oligosaccharide biosynthesis cooperatively but not in a redundant manner with beta-1,4-GalT I within cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / enzymology
  • Animals
  • Base Sequence
  • Blotting, Western
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cell Line
  • Colorectal Neoplasms / enzymology
  • DNA Primers
  • DNA, Complementary
  • Galactose / metabolism*
  • Galactosyltransferases / genetics
  • Galactosyltransferases / metabolism*
  • Humans
  • Isoenzymes / genetics
  • Isoenzymes / metabolism*
  • K562 Cells
  • Molecular Sequence Data
  • Oligosaccharides / chemistry
  • Oligosaccharides / metabolism*
  • Spodoptera


  • DNA Primers
  • DNA, Complementary
  • Isoenzymes
  • Oligosaccharides
  • Galactosyltransferases
  • Galactose

Associated data

  • GENBANK/AB024434
  • GENBANK/AB024435
  • GENBANK/AB024436