Glycobiology Advance Access originally published online on February 16, 2006
Glycobiology 2006 16(7):584-593; doi:10.1093/glycob/cwj090
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycosyltransferases involved in type 1 chain and Lewis antigen biosynthesis exhibit glycan and core chain specificity
Division of Clinical Immunology, Karolinska Institutet, Karolinska University Hospital, Huddinge, S-141 86 Stockholm, Sweden
1 To whom correspondence should be addressed; e-mail: jonas.lofling{at}ki.se
Received on October 19, 2005; revised on January 30, 2006; accepted on February 10, 2006
Sialyl Lewis A (SLea), Lewis A (Lea), and Lewis B (Leb) have been studied in many different biological contexts, for example in microbial adhesion and cancer. Their biosynthesis is complex and involves ß1,3-galactosyltransferases (ß3Gal-Ts) and a combined action of 
2- and/or 4-fucosyltransferases (Fuc-Ts). Further, O-glycans with different core structures have been identified, and the ability of ß3Gal-Ts and Fuc-Ts to use these as substrates has not been resolved. Therefore, to examine the in vivo specificity of enzymes involved in SLea, Lea, and Leb synthesis, we have transiently transfected CHO-K1 cells with relevant human glycosyltransferases and, on secreted reporter proteins, detected the resulting Lewis antigens on N- and O-linked glycans using western blotting and Le-specific antibodies. ß3Gal-T1, -T2, and -T5 could synthesize type 1 chains on N-linked glycans, but only ß3Gal-T5 worked on O-linked glycans. The latter enzyme could use both core 2 and core 3 precursor structures. Furthermore, the specificity of FUT5 and FUT3 in Lea and Leb synthesis was different, with FUT5 fucosylating H type 1 only on core 2, but FUT3 fucosylating H type 1 much more efficient on core 3 than on core 2. Finally, FUT1 and FUT2 were both found to direct 2-fucosylation on type 1 chains on both N- and O-linked structures. This knowledge enables us to engineer recombinant glycoproteins with glycan- and core chain-specific Lewis antigen substitution. Such tools will be important for investigations on the fine carbohydrate specificity of Leb-binding lectins, such as Helicobacter pylori adhesins and DC-SIGN, and may also prove useful as therapeutics.
Key words: blood group antigens / cancer associated epitopes / fucosyltransferase / Lewis antigens / ß3-galactosyltransferase
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Lofling, M. Diswall, S. Eriksson, T. Boren, M. E Breimer, and J. Holgersson Studies of Lewis antigens and H. pylori adhesion in CHO cell lines engineered to express Lewis b determinants Glycobiology, July 1, 2008; 18(7): 494 - 501. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Altman, H. Fernandez, V. Chandan, B. A. Harrison, M. W. Schuster, L. O. Rademacher, and C. Toledo Analysis of Helicobacter pylori isolates from Chile: occurrence of selective type 1 Lewis b antigen expression in lipopolysaccharide J. Med. Microbiol., May 1, 2008; 57(5): 585 - 591. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Strasser, J. S. Bondili, U. Vavra, J. Schoberer, B. Svoboda, J. Glossl, R. Leonard, J. Stadlmann, F. Altmann, H. Steinkellner, et al. A Unique {beta}1,3-Galactosyltransferase Is Indispensable for the Biosynthesis of N-Glycans Containing Lewis a Structures in Arabidopsis thaliana PLANT CELL, July 1, 2007; 19(7): 2278 - 2292. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Ma, J. L. Simala-Grant, and D. E. Taylor Fucosylation in prokaryotes and eukaryotes Glycobiology, December 1, 2006; 16(12): 158R - 184R. [Abstract] [Full Text] [PDF]
|
|