Glycobiology, 2000, Vol. 10, No. 8 837-847
© 2000 Oxford University Press
N-glycan patterns of human transferrin produced in Trichoplusia ni insect cells: effects of mammalian galactosyltransferase
Department of Chemical Engineering, The Johns Hopkins University, Baltimore, MD 21218, USA, 2GlycoLab, Central Research Institute, Nakano Vinegar Co. Ltd., 2–6 Nakamura-cho, Handa-City Aichi 475–8585, Japan, 3Suntory Institute for Bioorganic Research, Wakayama-dai, Shimamoto-cho, Mishima-gun, Osaka 618, Japan, the 4Department of Microbiology, University of Wyoming, Laramie, WY 82071, USA, and 5Department of Biology, The Johns Hopkins University, Baltimore, MD 21218, USA
The N-glycans of human serum transferrin produced in Trichopulsia ni cells were analyzed to examine N-linked oligosaccharide processing in insect cells. Metabolic radiolabeling of the intra- and extracellular protein fractions revealed the presence of multiple transferrin glycoforms with molecular weights lower than that observed for native human transferrin. Consequently, the N-glycan structures of transferrin in the culture medium were determined using three-dimensional high performance liquid chromatography. The attached oligosaccharides included high mannose, paucimannosidic, and hybrid structures with over 50% of these structures containing one fucose, 
(1,6)-, or two fucoses, (1,6)- and (1,3)-, linked to the Asn-linked N-acetylglucosamine. Neither sialic acid nor galactose was detected on any of the N-glycans. However, when transferrin was coexpressed with ß(1,4)-galactosyltransferase three additional galactose-containing hybrid oligosaccharides were obtained. The galactose attachments were exclusive to the (1,3)-mannose branch and the structures varied by the presence of zero, one, or two attached fucose residues. Furthermore, the presence of the galactosyltransferase appeared to reduce the number of paucimannosidic structures, which suggests that galactose attachment inhibits the ability of hexosaminidase activity to remove the terminal N-acetylglucosamine. The ability to promote galactosylation and reduce paucimannosidic N-glycans suggests that the oligosaccharide processing pathway in insect cells may be manipulated to mimic more closely that of mammalian cells.
1 To whom correspondence should be addressed
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Tomiya, S. Narang, J. Park, B. Abdul-Rahman, O. Choi, S. Singh, J. Hiratake, K. Sakata, M. J. Betenbaugh, K. B. Palter, et al. Purification, Characterization, and Cloning of a Spodoptera frugiperda Sf9 beta-N-Acetylhexosaminidase That Hydrolyzes Terminal N-Acetylglucosamine on the N-Glycan Core J. Biol. Chem., July 14, 2006; 281(28): 19545 - 19560. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Leonard, D. Rendic, C. Rabouille, I. B. H. Wilson, T. Preat, and F. Altmann The Drosophila fused lobes Gene Encodes an N-Acetylglucosaminidase Involved in N-Glycan Processing J. Biol. Chem., February 24, 2006; 281(8): 4867 - 4875. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Choi, N. Tomiya, J. H. Kim, J. M. Slavicek, M. J. Betenbaugh, and Y. C. Lee N-glycan structures of human transferrin produced by Lymantria dispar (gypsy moth) cells using the LdMNPV expression system Glycobiology, July 1, 2003; 13(7): 539 - 548. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kim, S. K. Hwang, R. A. Dwek, P. M. Rudd, Y. H. Ahn, E.-H. Kim, C. Cheong, S. I. Kim, N. S. Park, and S. M. Lee Structural determination of the N-glycans of a lepidopteran arylphorin reveals the presence of a monoglucosylated oligosaccharide in the storage protein Glycobiology, March 1, 2003; 13(3): 147 - 157. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Tomiya, D. Howe, J. J. Aumiller, M. Pathak, J. Park, K. B. Palter, D. L. Jarvis, M. J. Betenbaugh, and Y. C. Lee Complex-type biantennary N-glycans of recombinant human transferrin from Trichoplusia ni insect cells expressing mammalian {beta}-1,4-galactosyltransferase and {beta}-1,2-N-acetylglucosaminyltransferase II Glycobiology, January 1, 2003; 13(1): 23 - 34. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Marchal, M. Cerutti, A.-M. Mir, S. Juliant, G. Devauchelle, R. Cacan, and A. Verbert Expression of a membrane-bound form of Trypanosoma cruzi trans-sialidase in baculovirus-infected insect cells: a potential tool for sialylation of glycoproteins produced in the baculovirus-insect cells system Glycobiology, July 1, 2001; 11(7): 593 - 603. [Abstract] [Full Text] [PDF]
|
|