Introduction of H-antigens into oligosaccharides and sugar chains of glycoproteins using highly efficient 1,2-α-l-fucosynthase.

Publisher: IRL Press at Oxford University Press Country of Publication: England NLM ID: 9104124 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1460-2423 (Electronic) Linking ISSN: 09596658 NLM ISO Abbreviation: Glycobiology Subsets: MEDLINE

Original Publication: Oxford ; New York : IRL Press at Oxford University Press, c1990-

Antigens, Bacterial/*metabolism ; Glycoproteins/*metabolism ; Oligosaccharides/*metabolism ; Sugars/*metabolism ; alpha-L-Fucosidase/*metabolism; Antigens, Bacterial/chemistry ; Bifidobacterium bifidum/enzymology ; Biocatalysis ; Carbohydrate Conformation ; Glycoproteins/chemistry ; Models, Molecular ; Oligosaccharides/chemistry ; Sugars/chemistry

Fucα1-2 Gal linkages, or H-antigens, constitute histo-blood group antigens and are involved in various physiological processes. In addition, recent studies have shown that the H-antigen-containing glycans play an important role, not only in establishing harmonious relationship between gut microbes and the host, but also in preventing gut dysbiosis-related diseases. Therefore, development of an efficient method for introducing Fuc residue at Gal residue at the nonreducing end of glycans via α-(1→2) linkage is desired for research as well as medicinal purposes. In this study, we succeeded in derivatizing inverting 1,2-α-l-fucosidase (AfcA) into a highly efficient 1,2-α-l-fucosynthase. The synthase specifically synthesized H type 1-, type 2-, type 3- and type 4-chain-containing oligosaccharides with yields of 57-75% based on acceptor depletion. The synthase was also able to specifically introduce Fuc residues into Lewis a/x antigens to produce Lewis b/y antigens, with yields of 43% and 62%, respectively. In addition, the enzyme efficiently introduced H-antigens into sugar chains of porcine gastric mucins, as revealed by lectin blotting and mass spectroscopy analysis of the sugars. Detailed acceptor specificity analysis using various monosaccharides and oligosaccharides unraveled unique substrate recognition feature of this synthase at the subsite (+1), which can be explained by our previous X-ray crystallographic study of AfcA. These results show that the synthase developed in this study could serve as an alternative to other H-antigen synthesis methods involving α-1,2-fucosyltransferases and retaining α-fucosidase.
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Keywords: H-antigen; glycoprotein; glycosylation; glycosynthase; oligosaccharide

0 (Antigens, Bacterial)
0 (Glycoproteins)
0 (H antigen)
0 (Oligosaccharides)
0 (Sugars)
EC 3.2.1.51 (alpha-L-Fucosidase)

Date Created: 20160824 Date Completed: 20180405 Latest Revision: 20180412

20231215

10.1093/glycob/cww085

27550195