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Identification of a New alpha1,2-fucosyltransferase Involved in O-antigen Biosynthesis of Escherichia Coli O86:B7 and Formation of H-type 3 Blood Group Antigen

Identification of a New alpha1,2-fucosyltransferase Involved in O-antigen Biosynthesis of Escherichia Coli O86:B7 and Formation of H-type 3 Blood Group Antigen

Mei Li, Jie Shen, Xianwei Liu, Jun Shao, Wen Yi, Christine S Chow, Peng G Wang

Biochemistry. 2008 Nov 4;47(44):11590-7.

PMID: 18842005

Abstract:

Escherichia coli O86 possesses high human blood group B activity because of its O-antigen structure, sharing the human blood group B epitope. In this study, the wbwK gene of E. coli O86:B7 was expressed and purified as the GST fusion protein. Thereafter, the wbwK gene was biochemically identified to encode an alpha1,2-fucosyltransferase through radioactivity assays, as well as mass spectrometry and NMR spectroscopy. WbwK shows strict substrate specificity and only recognizes Gal beta1,3GalNAc alpha-OR (T-antigen and derivatives) as the acceptor to generate the H-type 3 blood group antigen. In contrast to other alpha1,2-fucosyltransferases, WbwK does not display activity toward the simple substrate Gal beta-OMe. Comparison with another recently characterized alpha1,2-fucosyltransferase (WbsJ) of E. coli O128:B12 indicates a low level of amino acid identity between them; however, they share a common acceptor substrate, Gal beta1,3GalNAc alpha-OR. Domain swapping between WbwK and WbsJ revealed that the smaller variable domains located in the C-terminus determine substrate specificity, whereas the larger variable domain in the N-terminus might play a role in forming the correct conformation for substrate binding or for localization of the alpha1,2-fucosyltransferase involved in O-antigen biosynthesis. In addition, milligram scale biosynthesis of the H-type 3 blood group antigen was explored using purified recombinant WbwK. WbwK may have potential applications in masking T-antigen, the tumor antigen, in vivo.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
LS741397	Lipopolysaccharides from Escherichia coli O128:B12			Price

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