The Acceptor and Site Specificity of α3-Fucosyltransferase V

Pykäri, Maria; Toivonen, Sanna; Natunen, Jari; Niemelä, Ritva; Salminen, Heidi; Aitio, Olli; Ekström, Minna; Parmanne, Pinja; Välimäki, Mika J.; Alais, Jocelyne; Augé, Claudine; Lowe, John B.; Renkonen, Ossi; Renkonen, Risto

doi:10.1074/jbc.m007922200

Cited by 13 publications

(4 citation statements)

References 65 publications

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“…Previous studies have demonstrated that cells transfected with full-length FucT-V and FucT-VI express a different complement of cell-surface fucosylated antigens (14,26). Cells transfected with FucT-V preferentially produce VIM2, whereas cells transfected with FucT-VI preferentially produce sialyl-Le x ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…One such subtle difference is the site-specific fucosylation properties of FucT-V and FucT-VI with fucose transfer to sialylated polylactosamine structures. FucT-V has been shown to favor fucosylation of internal GlcNAc residues, yielding the VIM2 determinant structure, whereas FucT-VI preferentially fucosylates distal GlcNAc residues, leading to the sialyl-Le x determinant structure (14,26). The difference in the fucosylation specificity of the two FucTs leads to the production of products that have different biological functions and antigenicity (38,39).…”

Section: Discussionmentioning

confidence: 99%

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Site-specific Fucosylation of Sialylated Polylactosamines by α1,3/4-Fucosyltransferases-V and -VI Is Defined by Amino Acids Near the N Terminus of the Catalytic Domain

Shetterly

Jost

Watson

et al. 2007

Journal of Biological Chemistry

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Fucose transfer from GDP-fucose to GlcNAc residues of the sialylated polylactosamine acceptor NeuAc␣2-3Gal␤1-4Glc-NAc␤1-3Gal␤1-4GlcNAc␤1-3Gal␤1-4Glc␤1-ceramide leads to two isomeric monofucosyl antigens, VIM2 and sialyl-Le x . Human ␣1,3/4-fucosyltransferase (FucT)-V catalyzes primarily the synthesis of VIM2, whereas human FucT-VI catalyzes primarily the synthesis of sialyl-Le x . Thus, these two enzymes have distinct "site-specific fucosylation" properties. Amino acid sequence alignment of these enzymes showed that there are 24 amino acid differences in their catalytic domains. Studies were conducted to determine which of the amino acid differences are responsible for the site-specific fucosylation properties of each enzyme. Domain swapping (replacing a portion of the catalytic domain from one enzyme with an analogous portion from the other enzyme) demonstrated that site-specific fucosylation was defined within a 40-amino acid segment containing 8 amino acid differences between the two enzymes. Site-directed mutagenesis studies demonstrated that the site-specific fucosylation properties of these enzymes could be reversed by substituting 4 amino acids from one sequence with the other. These results were observed in both in vitro enzyme assays and flow cytometric analyses of Chinese hamster ovary cells transfected with plasmids containing the various enzyme constructs. Modeling studies of human FucT using a structure of a bacterial fucosyltransferase as a template demonstrated that the amino acids responsible for site-specific fucosylation map near the GDP-fucose-binding site. Additional enzyme studies demonstrated that FucT-VI has ϳ12-fold higher activity compared with FucT-V and that the Trp 124 /Arg 110 site in these enzymes is responsible primarily for this activity difference. ␣1,3/4-Fucosyltransferases (FucTs)3 catalyze transfer of fucose to GlcNAc residues present in lacto or neolacto series structures of cell-surface glycolipids and glycoproteins. Fucosylated structures are known to accumulate in many human cancers (1-4), function as ligands for leukocyte adhesion during inflammation (4, 5), and undergo developmental regulation (6 -8). The diversity of naturally occurring ␣1,3/4-fucosylated structures found in human cells is controlled by a family of six distinct yet related FucTs with individual tissue distribution properties and acceptor substrate preferences. Among these six FucTs, FucT-III, FucT-V, and FucT-VI share Ͼ85% amino acid sequence homology and originated from a common ancestral gene via gene duplication (9 -11). Genes for these enzymes form a cluster on chromosome 19 in humans (12-15). FucT-IV, FucT-VII, and FucT-IX share a lower amino acid sequence homology both between each other and with FucT-III, FucT-V, and FucT-VI. Additionally, they have distinct chromosomal localization. Genes for FucT-IV, FucT-VII, and FucT-IX have been mapped to chromosomes 11, 9, and 6, respectively (16 -18).Although all of these enzymes catalyze fucose transfer to (LacNAc)Gal␤1-4GlcNAc chain acceptors to produce ...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Site-specific Fucosylation of Sialylated Polylactosamines by α1,3/4-Fucosyltransferases-V and -VI Is Defined by Amino Acids Near the N Terminus of the Catalytic Domain

Shetterly

Jost

Watson

et al. 2007

Journal of Biological Chemistry

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“…Mammalian sperm express fucosyltransferase activities, whereas FUT5 is found only in humans and chimpanzees [58]. FUT5 is thought to recognize N -acetyllactosamines (Galβ1-3GlcNAc, Galβ1-4GlcNAc) as well as GalNAcβ1-4GlcNAc, regardless of their modification by sulfation, sialylation, or fucosylation [59,60]. FUT5 is an integral membrane protein with an intracellularly oriented N-terminus and extracellularly oriented C-terminus.…”

Section: Sperm Recognition Of the Zpmentioning

confidence: 99%

Protein-Carbohydrate Interaction between Sperm and the Egg-Coating Envelope and Its Regulation by Dicalcin, a Xenopus laevis Zona Pellucida Protein-Associated Protein

Miwa

2015

Molecules

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Protein-carbohydrate interaction regulates multiple important processes during fertilization, an essential biological event where individual gametes undergo intercellular recognition to fuse and generate a zygote. In the mammalian female reproductive tract, sperm temporarily adhere to the oviductal epithelium via the complementary interaction between carbohydrate-binding proteins on the sperm membrane and carbohydrates on the oviductal cells. After detachment from the oviductal epithelium at the appropriate time point following ovulation, sperm migrate and occasionally bind to the extracellular matrix, called the zona pellucida (ZP), which surrounds the egg, thereafter undergoing the exocytotic acrosomal reaction to penetrate the envelope and to reach the egg plasma membrane. This sperm-ZP interaction also involves the direct interaction between sperm carbohydrate-binding proteins and carbohydrates within the ZP, most of which have been conserved across divergent species from mammals to amphibians and echinoderms. This review focuses on the carbohydrate-mediated interaction of sperm with the female reproductive tract, mainly the interaction between sperm and the ZP, and introduces the fertilization-suppressive action of dicalcin, a Xenopus laevis ZP protein-associated protein.The action of dicalcin correlates significantly with a dicalcin-dependent change in the lectin-staining pattern within the ZP, suggesting a unique role of dicalcin as an inherent protein that is capable of regulating the affinity between the lectin and oligosaccharides attached on its target glycoprotein.

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“…Neu5Ac␣2-3Gal␤1-4GlcNAc␤1-3Gal␤1-4GlcNAc was enzymatically synthesized as described in Ref. 17. UDP-GalNAc, CMP-Neu5Ac, bovine milk ␤1,4-galactosyltransferase, and jack bean ␤-N-acetylhexosaminidase were from Sigma.…”

Section: Methodsmentioning

confidence: 99%

α2,3-Sialylation of Terminal GalNAcβ1–3Gal Determinants by ST3Gal II Reveals the Multifunctionality of the Enzyme

Toivonen¹,

Aitio²,

Renkonen³

2001

Journal of Biological Chemistry

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Enzymatic ␣2,3-sialylation of GalNAc has not been described previously, although some glycoconjugates containing ␣2,3-sialylated GalNAc residues have been reported. In the present experiments, recombinant soluble ␣2,3-sialyltransferase ST3Gal II efficiently sialylated the X 2 pentasaccharide GalNAc␤1-3Gal␤1-4GlcNAc␤1-3Gal␤1-4Glc, globo-N-tetraose GalNAc␤1-3Gal␣1-4Gal␤1-4Glc, and the disaccharide GalNAc␤1-3Gal in vitro. The purified products were identified as Neu5Ac␣2-3GalNAc␤1-3Gal␤1-4GlcNAc␤1-3Gal␤1-4Glc, Neu5Ac␣2-3GalNAc␤1-3Gal␣1-4Gal␤1-4Glc, and Neu5Ac␣2-3GalNAc␤1-3Gal, respectively, by matrixassisted laser desorption/ionization time-of-flight mass spectrometry, enzymatic degradations, and oneand two-dimensional NMR-spectroscopy. In particular, the presence of the Neu5Ac␣2-3GalNAc linkage was firmly established in all three products by a long range correlation between Neu5Ac C2 and GalNAc H3 in heteronuclear multiple bond correlation spectra. Collectively, the data describe the first successful sialyltransfer reactions to the 3-position of GalNAc in any acceptor. Previously, ST3Gal II has been shown to transfer to the Gal␤1-3GalNAc determinant. Consequently, the present data show that the enzyme is multifunctional, and could be renamed ST3Gal(NAc) II. In contrast to ST3Gal II, ST3Gal III did not transfer to the X 2 pentasaccharide. The Neu5Ac␣2-3GalNAc linkage of sialyl X 2 was cleaved by sialidases from Arthrobacter ureafaciens and Clostridium perfringens, but resisted the action of sialidases from Newcastle disease virus and Streptococcus pneumoniae. Therefore, the latter two enzymes cannot be used to differentiate between Neu5Ac␣2-3GalNAc and Neu5Ac␣2-6GalNAc linkages, as has been assumed previously.Enzymatic ␣2,3-sialylation of GalNAc 1 has not been described before, although structures containing ␣2,3-sialylated GalNAc residues, like sialyl X 2 , have been reported. The X 2 epitope, GalNAc␤1-3Gal␤1-4GlcNAc␤1-3Gal␤1-4Glc, and its distally ␣2,3-sialylated form have been characterized from glycosphingolipids on human red blood cells (1-3). Immunohistochemical experiments with the anti-X 2 monoclonal antibody TH2 suggest the presence of X 2 and sialylated X 2 also in human white blood cells, liver, spleen, lung, kidney, colon, pancreas, brain, and placenta (3). High concentration of X 2 glycolipid has been reported in gastric tumor tissue (4). Sialyl globoside (Neu5Ac␣2-3GalNAc␤1-3Gal␣1-4Gal␤1-4Glc) has been characterized from human teratocarcinoma cells (5) and from muscles affected by amyotrophic lateral sclerosis (6) by chromatographic behavior, exoglycosidase treatments, and antibody reactivity. Additionally, sialyl LacdiNAc (Neu5Ac␣2-3GalNAc␤1-4GlcNAc) from snake venom serine proteases contains ␣2,3-sialylated GalNAc (7, 8). It has been suggested that the X 2 epitope is the human receptor for Clostridium difficile toxin A (9). The X 2 structure also occurs in the lipooligosaccharide of Neisseria gonorrhoeae strain F62 (10).The known mammalian ␣2,3-sialyltransferases, ST3Gal I-VI, 2 transfer sialic acid...

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The Acceptor and Site Specificity of α3-Fucosyltransferase V

Cited by 13 publications

References 65 publications

Site-specific Fucosylation of Sialylated Polylactosamines by α1,3/4-Fucosyltransferases-V and -VI Is Defined by Amino Acids Near the N Terminus of the Catalytic Domain

Site-specific Fucosylation of Sialylated Polylactosamines by α1,3/4-Fucosyltransferases-V and -VI Is Defined by Amino Acids Near the N Terminus of the Catalytic Domain

Protein-Carbohydrate Interaction between Sperm and the Egg-Coating Envelope and Its Regulation by Dicalcin, a Xenopus laevis Zona Pellucida Protein-Associated Protein

α2,3-Sialylation of Terminal GalNAcβ1–3Gal Determinants by ST3Gal II Reveals the Multifunctionality of the Enzyme

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