3UET

Crystal structure of alpha-1,3/4-fucosidase from Bifidobacterium longum subsp. infantis D172A/E217A mutant complexed with lacto-N-fucopentaose II

Summary for 3UET

• Entry DOI: 10.2210/pdb3uet/pdb
• Related: 3MO4 3UES
• Related PRD ID: PRD_900129
• Descriptor: Alpha-1,3/4-fucosidase, beta-D-galactopyranose-(1-3)-[alpha-L-fucopyranose-(1-4)]2-acetamido-2-deoxy-beta-D-glucopyranose, SODIUM ION, ... (5 entities in total)
• Functional Keywords: tim barrel, hydrolase
• Biological source: Bifidobacterium longum subsp. infantis
• Total number of polymer chains: 2
• Total formula weight: 107210.90

Authors

Sakurama, H.,Fushinobu, S.,Yoshida, E.,Honda, Y.,Hidaka, M.,Ashida, H.,Kitaoka, M.,Katayama, T.,Yamamoto, K.,Kumagai, H. (deposition date: 2011-10-31, release date: 2012-04-04, Last modification date: 2023-11-01)

Primary citation

Sakurama, H.,Fushinobu, S.,Hidaka, M.,Yoshida, E.,Honda, Y.,Ashida, H.,Kitaoka, M.,Kumagai, H.,Yamamoto, K.,Katayama, T.
1,3-1,4-alpha-L-fucosynthase that specifically introduces Lewis a/x antigens into type-1/2 chains
J.Biol.Chem., 287:16709-16719, 2012

PubMed Abstract: α-L-fucosyl residues attached at the non-reducing ends of glycoconjugates constitute histo-blood group antigens Lewis (Le) and ABO and play fundamental roles in various biological processes. Therefore, establishing a method for synthesizing the antigens is important for functional glycomics studies. However, regiospecific synthesis of glycosyl linkages, especially α-L-fucosyl linkages, is quite difficult to control both by chemists and enzymologists. Here, we generated an α-L-fucosynthase that specifically introduces Le(a) and Le(x) antigens into the type-1 and type-2 chains, respectively; i.e. the enzyme specifically accepts the disaccharide structures (Galβ1-3/4GlcNAc) at the non-reducing ends and attaches a Fuc residue via an α-(1,4/3)-linkage to the GlcNAc. X-ray crystallographic studies revealed the structural basis of this strict regio- and acceptor specificity, which includes the induced fit movement of the catalytically important residues, and the difference between the active site structures of 1,3-1,4-α-L-fucosidase (EC 3.2.1.111) and α-L-fucosidase (EC 3.2.1.51) in glycoside hydrolase family 29. The glycosynthase developed in this study should serve as a potentially powerful tool to specifically introduce the Le(a/x) epitopes onto labile glycoconjugates including glycoproteins. Mining glycosidases with strict specificity may represent the most efficient route to the specific synthesis of glycosidic bonds.

PubMed: 22451675
DOI: 10.1074/jbc.M111.333781

Experimental method

X-RAY DIFFRACTION (2.1 Å)