Human α1,3/4 Fucosyltransferases

Human α1,3 fucosyltransferases (FucTs) contain four highly conserved cysteine (Cys) residues, in addition to a free Cys residue that lies near the binding site for GDP-fucose (Holmes, E. H., Xu, Z., Sherwood, A. L., and Macher, B. A. (1995)J. Biol. Chem. 270, 8145–8151). The participation of the highly conserved Cys residues in disulfide bonds and their functional significance were characterized by mass spectrometry (MS) analyses and site-directed mutagenesis, respectively. Among the human FucTs is a subset of enzymes (FucT III, V, and VI) having highly homologous sequences, especially in the catalytic domain, and Cys residues in FucT III and V were characterized. The amino acid sequence of FucT III was characterized. Peptides containing the four conserved Cys residues were detected after reduction and alkylation, and found to be involved in disulfide bonds. The disulfide bond pattern was characterized by multiple stage MS analysis and the use of Glu-C protease and MS/MS analysis. Disulfide bonds in FucT III occur between Cys residues (Cys81 to Cys338 and Cys91 to Cys341) at the N and C termini of the catalytic domain, bringing these ends close together in space. Mutagenesis of highly conserved Cys residues to Ser in FucT V resulted in proteins lacking enzymatic activity. Three of the four mutants have molecular weights similar to wild type enzyme and maintained an ability to bind GDP, whereas the other (Cys104) produced a series of lower molecular weight bands when characterized by Western blot analysis, and did not bind GDP. FucTs have highly conserved, potentialN-linked sites, and our mass spectrometry analyses demonstrated that both N-linked sites are modified with oligosaccharides.

• Publication year

  2000

• Venue

  Journal of Biological Chemistry

• Publication date

  2000-08-11

• Fields of study

  Biology, Chemistry

• Identifiers
  DOI 10.1074/jbc.M000888200
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

• α1,3‐Fucoslytransferase IX (Fuc‐TIX) is very highly conserved between human and mouse; molecular cloning, characterization and tissue distribution of human Fuc‐TIX

  1999cited by this paper
• Molecular Cloning and Characterization of Two Zebrafish α(l,3)Fucosyltransferase Genes Developmentally Regulated in Embryogenesisxy

  1999cited by this paper
• A Single Amino Acid in the Hypervariable Stem Domain of Vertebrate α1,3/1,4-Fucosyltransferases Determines the Type 1/Type 2 Transfer

  1999cited by this paper
• Divergent evolution of fucosyltransferase genes from vertebrates, invertebrates, and bacteria.

  1999cited by this paper
• Purification, cDNA Cloning, and Expression of GDP-l-Fuc:Asn-linked GlcNAc α1,3-Fucosyltransferase from Mung Beans*

  1999cited by this paper
• Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris.

  1998cited by this paper
• Conserved structural features in eukaryotic and prokaryotic fucosyltransferases.

  1998cited by this paper
• Molecular cloning and characterization of an alpha1,3 fucosyltransferase, CEFT-1, from Caenorhabditis elegans.

  1998cited by this paper
• Human α1,3/4-Fucosyltransferases

  1998cited by this paper
• Human α1,3/4-Fucosyltransferases

  1998cited by this paper
• Human α1,3/4-Fucosyltransferases

  1998cited by this paper
• Molecular Cloning and Expression of a Bovine α(1,3)-Fucosyltransferase Gene Homologous to a Putative Ancestor Gene of the Human FUT3-FUT5-FUT6 Cluster*

  1997cited by this paper
• Structure-function analysis of human alpha1,3-fucosyltransferase. Amino acids involved in acceptor substrate specificity.

  1996cited by this paper
• Human α(1,3/1,4)-Fucosyltransferases Discriminate between Different Oligosaccharide Acceptor Substrates through a Discrete Peptide Fragment (*)

  1995cited by this paper
• Structure-function analysis of human alpha 1-->3fucosyltransferases. A GDP-fucose-protected, N-ethylmaleimide-sensitive site in FucT-III and FucT-V corresponds to Ser178 in FucT-IV.

  1995cited by this paper
• Acceptor specificity of different length constructs of human recombinant alpha 1,3/4-fucosyltransferases. Replacement of the stem region and the transmembrane domain of fucosyltransferase V by protein A results in an enzyme with GDP-fucose hydrolyzing activity.

  1995cited by this paper
• Expression cloning of a novel alpha 1,3-fucosyltransferase that is involved in biosynthesis of the sialyl Lewis x carbohydrate determinants in leukocytes.

  1994cited by this paper
• Molecular cloning of a cDNA encoding a novel human leukocyte alpha-1,3-fucosyltransferase capable of synthesizing the sialyl Lewis x determinant.

  1994cited by this paper
• Tandem mass spectrometry of very large molecules. 2. Dissociation of multiply charged proline-containing proteins from electrospray ionization.

  1993cited by this paper
• Molecular cloning of a fourth member of a human alpha (1,3)fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis x, sialyl Lewis x, and difucosyl sialyl Lewis x epitopes.

  1992cited by this paper
• Molecular cloning of a human fucosyltransferase gene that determines expression of the Lewis x and VIM-2 epitopes but not ELAM-1-dependent cell adhesion.

  1991cited by this paper
• Human alpha 1-3 fucosyltransferases.

  1991cited by this paper
• Photoaffinity labeling of GDP-fucose:nLcOse4Cer alpha 1----3-fucosyltransferase from human small cell lung carcinoma NCI-H69 cells with the GDP-fucose analog GDP-hexanolaminyl-4-azidosalicylic acid.

  1990cited by this paper
• PCR protocols — A guide to methods and applications

  1990cited by this paper

• Characterization of MiFUT11 from Mangifera indica L.: A functional core α1,3-fucosyltransferase potentially involved in the biosynthesis of immunogenic carbohydrates in mango fruit.

  2019cites this paper
• Characterization of a GDP-Fucose Transporter and a Fucosyltransferase Involved in the Fucosylation of Glycoproteins in the Diatom Phaeodactylum tricornutum

  2019cites this paper
• Molecular cloning and functional expression of Lewis type α1,3/α1,4-fucosyltransferase cDNAs from Mangifera indica L.

  2017cites this paper
• Glycosyltransferases: mechanisms and applications in natural product development.

  2015cites this paper
• Exploring the N-glycosylation Pathway in Chlamydomonas reinhardtii Unravels Novel Complex Structures*

  2013cites this paper
• NOVEL COMPLEX STRUCTURES.

  2013cites this paper
• N-Glycans of Phaeodactylum tricornutum Diatom and Functional Characterization of Its N-Acetylglucosaminyltransferase I Enzyme*

  2010cites this paper
• Human fucosyltransferase IX: Production, purification and characterisation

  2010cites this paper
• Structure – function study of Arabidopsis thaliana core alpha1,3-fucosyltransferase (FucTA)

  2009cites this paper
• Down-regulated expression of plant-specific glycoepitopes in alfalfa.

  2008cites this paper
• 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*

  2007cites this paper
• Human xylosyltransferases in health and disease

  2007cites this paper
• GDP-L-fucose: synthesis and role in inflammation

  2007cites this paper
• Construction of a Library of Human Glycosyltransferases Immobilized in the Cell Wall of Saccharomyces cerevisiae

  2006cites this paper
• Fucosylation in prokaryotes and eukaryotes.

  2006cites this paper
• Purification, Kinetic Characterization, and Mapping of the Minimal Catalytic Domain and the Key Polar Groups of Helicobacter pylori α-(1,3/1,4)-Fucosyltransferases*

  2006cites this paper
• Expression und Charakterisierung der humanen Fucosyltransferase V

  2006cites this paper
• High level expression of monomeric and dimeric human α1,3-fucosyltransferase V

  2006cites this paper
• Determination of glycosylation sites and disulfide bond structures using LC/ESI-MS/MS analysis.

  2006cites this paper
• Human xylosyltransferase I: functional and biochemical characterization of cysteine residues required for enzymic activity.

  2005cites this paper
• Untersuchungen zur biochemischen Charakterisierung der humanen Xylosyltransferase I und II

  2005cites this paper
• Structure/function study of Lewis alpha3- and alpha3/4-fucosyltransferases: the alpha1,4 fucosylation requires an aromatic residue in the acceptor-binding domain.

  2004cites this paper
• Identification of Disulfide Bonds among the Nine Core 2 N-Acetylglucosaminyltransferase-M Cysteines Conserved in the Mucin β6-N-Acetylglucosaminyltransferase Family*

  2004cites this paper
• Structure/function study of Lewis a 3- and a 3/4-fucosyltransferases: the a 1,4 fucosylation requires an aromatic residue in the acceptor-binding domain

  2004cites this paper
• Analysis of the site-specific N-glycosylation of beta1,6 N-acetylglucosaminyltransferase V.

  2004cites this paper
• Highly Conserved Cysteines of Mouse Core 2 β1,6-N-Acetylglucosaminyltransferase I Form a Network of Disulfide Bonds and Include a Thiol That Affects Enzyme Activity*

  2003cites this paper
• Chemical modifications of alpha1,6-fucosyltransferase define amino acid residues of catalytic importance.

  2003cites this paper
• Soluble human core 2 beta6-N-acetylglucosaminyltransferase C2GnT1 requires its conserved cysteine residues for full activity.

  2003cites this paper
• Alpha1,4-fucosyltransferase activity: a significant function in the primate lineage has appeared twice independently.

  2002cites this paper
• A highly conserved His-His motif present in alpha1-->3/4fucosyltransferases is required for optimal activity and functions in acceptor binding.

  2002cites this paper
• Characterizing closely spaced, complex disulfide bond patterns in peptides and proteins by liquid chromatography/electrospray ionization tandem mass spectrometry.

  2002cites this paper
• The presence of Lewis a epitopes in Arabidopsis thaliana glycoconjugates depends on an active alpha4-fucosyltransferase gene.

  2002cites this paper
• Identification of a cDNA encoding a plant Lewis-type α1,4-fucosyltransferase

  2001cites this paper
• Unique Disulfide Bond Structures Found in ST8Sia IV Polysialyltransferase Are Required for Its Activity*

  2001cites this paper
• Neighboring cysteine residues in human fucosyltransferase VII are engaged in disulfide bridges, forming small loop structures.

  2001cites this paper
• MINI REVIEW Fucosyltransferases: structure/function studies

  2001cites this paper
• Fucosyltransferases: structure/function studies.

  2001influential citation
• Cloning and expression of cDNAs encoding alpha1,3-fucosyltransferase homologues from Arabidopsis thaliana.

  2001cites this paper
• Disulfide Bonds of GM2 Synthase Homodimers

  2000cites this paper
• Ancestral exonic organization of FUT8, the gene encoding the alpha6-fucosyltransferase, reveals successive peptide domains which suggest a particular three-dimensional core structure for the alpha6-fucosyltransferase family.

  2000cites this paper