Skip to main content
SpringerLink
Log in

Effect of the manganese ion on human α3/4 fucosyltransferase III activity

  • Published:
Biometals Aims and scope Submit manuscript

Abstract

The effect of manganese and other divalent cations on the activity of a soluble recombinant form of human α3/4 fucosyltransferase III (SFT3) expressed in Spodoptera frugiperda (Sf9) insect cells was studied. SFT3 was active in the absence of divalent cations with an optimum pH of 4.5. In the absence of Mn2+ increasing the pH from 4.5 to 7.0 caused a decrease in the affinity of SFT3 for the acceptor Galβ3GlcNAcO(CH2)3NHCO(CH2)5NH-biotin, as monitored by the 4-fold increase in the apparent KM value (0.9 to 3.3 mM). At pH 7.0, the addition of Mn2+ activated the enzyme and caused an increase in the affinity of SFT3 for the acceptor, as monitored by the 5-fold decrease of the apparent KM value (3.3 to 0.7 mM). In solution, a complex between GDP-Fuc donor and the divalent cation Mn2+ was observed by electrospray ionization mass spectrometry, in a 1:1 stoichiometry. These results indicated that Mn2+ bound the enzyme and increased its affinity for the acceptor; one possible functional role of manganese in catalysis could be as an electrophilic catalyst, co-ordinating the negative charges of the phosphate groups of the GDP-Fuc donor and promoting Fuc transfer. At low pH values such role would be played by the proton.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Altschul SF, Madden TL, Schaffer AA et al. 1997 Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.

    Google Scholar 

  • Ash DE, Schramm V L. 1982 Determination of free and bound manganese(II) in hepatocytes from fed and fasted rats. J Biol Chem 257, 9261–9264.

    Google Scholar 

  • Bensadoun A, Wienstein D. 1976 Assay of proteins in the presence of interfering materials. Anal Biochem 70, 241–250.

    Google Scholar 

  • Beyer TA, Hill RL. 1980 Enzymatic properties of the betagalactoside alpha 1 leads to 2 fucosyltransferase from porcine submaxillary gland. J Biol Chem 255, 5373–5379.

    Google Scholar 

  • Boix E, Swaminathan GJ, Zhang R, Natesh R, Brew K, Acharya KR. 2001 Structure of UDP complex of UDP-galactose:betagalactoside-alpha-1,3-galactosyltransferase at 1.53-Å resolution reveals a conformational change in the catalytically important C terminus. J Biol Chem 276, 48608–48614.

    Google Scholar 

  • Breton C, Oriol R, Imberty A. 1998 Conserved structural features in eukaryotic and prokaryotic fucosyltransferases. Glycobiology 8, 87–94.

    Google Scholar 

  • Breton C, Imberty A. 1999 Structure/function studies of glycosyltransferases. Curr Opin Struc Biol 9, 563–571.

    Google Scholar 

  • Cornish-Bowden A. 1995 Fundamentals of Enzyme Kinetics, London: Portland Press; 19–53.

    Google Scholar 

  • Costa J., Grabenhorst E., Nimtz M., Conradt H.S. 1997 Stable expression of the Golgi form and secretory variants of human fucosyltransferase III from BHK-21 cells. J Biol Chem 272, 11613–11621.

    Google Scholar 

  • De Vries T, Srnka CA, Palcic MM, Swiedler SJ, van den Eijnden DH, Macher BA. 1995 Acceptor specificity of different length constructs of human recombinant α 1,3/4-fucosyltransferases. J Biol Chem 270, 8712–8722.

    Google Scholar 

  • Dupuy F, Petit JM, Mollicone R, Oriol R, Julien R, Maftah A. 1999 A single amino acid in the hypervariable stem domain of vertebrate α 1,3/1,4-Fucosyltransferases determines the type 1/type 2 transfer. J Biol Chem 274, 12257–12262.

    Google Scholar 

  • FraÚsto da Silva JJR, Williams RJP. 1997 The biological chemistry of elements-the inorganic chemistry of life, Oxford: Oxford University Press Inc.

    Google Scholar 

  • Gastinel LN, Cambillau C, Bourne Y. 1999 Crystal structures of the bovine beta4galactosyltransferase catalytic domain and its complex with uridine diphosphogalactose. Embo J 18, 3546–3557.

    Google Scholar 

  • Gastinel LN, Bignon C., Misra AK, Hindsgaul O, Shaper JH, Joziasse DH. 2001 Bovine alpha1,3-galactosyltransferase catalytic domain structure and its relationship with ABO histo-blood group and glycosphingolipid glycosyltransferases. Embo J 20, 638–649.

    Google Scholar 

  • Holmes EH, Xu Z, Sherwood AL, Macher BA. 1995 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. J Biol Chem 270, 8145–8151.

    Google Scholar 

  • Huang MC, ZÖllner O, Moll T et al. 2000 P-selectin glycoprotein ligand-1 and E-selectin ligand-1 are differentially modified by fucosyltransferases Fuc-TIV and Fuc-TVII in mouse neutrophils. J Biol Chem 275, 31353–31360.

    Google Scholar 

  • Ilver D, Arnqvist A, Ögren J et al. 1998 Helicobacter pylori adhesin binding fucosylated histo-blood group antigens revealed by retagging. Science 279, 373–377.

    Google Scholar 

  • Johnson PH, Donald ASR, Feeney J, Watkins WM. 1992 Reassessment of the acceptor specificity and general properties of the Lewis blood-group gene associated α-3/4-fucosyltransferase purified from human milk. Glycoconj J 9, 251–264.

    Google Scholar 

  • Johnson PH, Donald ASR, Watkins WM 1993 Purification and properties of the alpha-3/4-L-fucosyltransferase released into the culture medium during the growth of the human A431 epidermoid carcinoma cell line. Glycoconj J 10, 152–164.

    Google Scholar 

  • Kaneko M, Kudo T, Iwasaki H et al. 1999 α 1,3-Fucosyltransferase IX (Fuc-TIX) is very highly conserved between human and mouse; molecular cloning, characterization and tissue distribution of human Fuc-TIX. FEBS Lett 452, 237–242.

    Google Scholar 

  • Kukowska-Latallo JF, Larsen RD, Nair RP, Lowe JB. 1990 A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group α(1,3/1,4)fucosyltransferase. Genes Dev 4, 1288–1303.

    Google Scholar 

  • Laemmli UK 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T. Nature 227, 680–685.

    Google Scholar 

  • Morais VA, Serpa J, Palma AS, Costa T, Maranga L, Costa J. 2001 Expression and characterization of recombinant human α-3/4-fucosyltransferase III from Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) cells using the baculovirus expression system. Biochem J 353, 719–725.

    Google Scholar 

  • Murray BW, Takayama S, Schultz J, Wong C-H. 1996 Mechanism and Specificity of Human α-1,3-Fucosyltransferase V. Biochemistry 35, 11183–11195.

    Google Scholar 

  • Pang H, Koda Y, Soejima M, Kimura H. 1998 Significance of each of three missense mutations, G484A, G667A and G808A, present in an inactive allele of the human Lewis gene (FUT3) for α (1,3/1,4) fucosyltranferase inactivation. Glycoconjugate J 15, 961–967.

    Google Scholar 

  • Pedersen LC, Tsuchida K, Kitagawa H, Sugahara K, Darden TA, Negishi M. 2000 Heparan/chondroitin sulfate biosynthesis. Structure and mechanism of human glucuronyltransferase I. J Biol Chem 275, 34580–34585.

    Google Scholar 

  • Shoreibah MG, Hindsgaul O, Pierce M. 1992 Purification and characterization of rat kidney UDP-N-acetylglucosamine:alpha-6-D-mannoside beta-1,6-N-acetylglucosaminyltransferase. J Biol Chem 267, 2920–2927.

    Google Scholar 

  • Sinnott, ML. 1991 Catalytic mechanisms of enzymic glycosyl transfer. Chem Rev 90, 1170–1202.

    Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT et al. 1985 Measurement of protein using Bicinchoninic Acid. Anal Biochem 150, 76–85.

    Google Scholar 

  • Sousa VL, Costa MT, Palma AS, Enguita F, Costa J. 2001 Localization, purification and specificity of the full-length membrane-bound form of human recombinant alpha1,3/4-fucosyltransferase from BHK-21B cells. Biochem J 357, 803–810.

    Google Scholar 

  • Takada A, Ohmori K, Takahashi N. et al. 1991 Adhesion of human cancer cells to vascular endothelium mediated by a carbohydrate antigen, Sialyl Lewis A. Biochem Biophys Res Commun 179, 713–719.

    Google Scholar 

  • Tsopanakis AD, Herries DG. 1978 Bovine galactosyl transferase. Substrate.manganese complexes and the role of manganese ions in the mechanism. Eur J Biochem 83, 179–188.

    Google Scholar 

  • Ñnligil UM, Zhou S, Yuwaraj S, Sarkar M, Schachter H, Rini JM. 2000 X-ray crystal structure of rabbit N-acetylglucosaminyltransferase I: Catalytic mechanism and a new protein superfamily. Embo J 19, 5269–5280.

    Google Scholar 

  • Weston BW, Nair RP, Larsen RD, Lowe JB. 1992 Isolation of a novel human α(1,3) fucosyltransferase gene and molecular comparison to the human lewis blood group α(1,3/1,4) Fucosyltransferase Gene. J Biol Chem 267, 4152–4160.

    Google Scholar 

  • Wiggins CAR, Munro S. 1998 Activity of the yeast MNN1 α-1,3-mannosyltransferase requires a motif conserved in many other families of glycosyltransferases. J Biol Chem 95, 7945–7950.

    Google Scholar 

  • Witsell DL, Casey CE, Neville MC. 1990 Divalent cation activation of galactosyltransferase in native mammary Golgi vesicles. J Biol Chem 265, 15731–15737.

    Google Scholar 

  • Zhang Y, Wang PG, Brew K. 2001 Specificity and mechanism of metal ion activation in UDP-galactose:beta-galactoside-alpha-1,3-galactosyltransferase. J Biol Chem 276, 11567–11574.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Instituto de Tecnologia Química e Biológica, Oeiras, Portugal

    Angelina S. Palma, Vanessa A. Morais, Ana V. Coelho & Júlia Costa

  2. Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal

    Angelina S. Palma & Vanessa A. Morais

  3. Chemistry Department, Universidade de Évora, Évora, Portugal

    Ana V. Coelho

Authors
  1. Angelina S. Palma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vanessa A. Morais
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ana V. Coelho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Júlia Costa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Júlia Costa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Palma, A.S., Morais, V.A., Coelho, A.V. et al. Effect of the manganese ion on human α3/4 fucosyltransferase III activity. Biometals 17, 35–43 (2004). https://doi.org/10.1023/A:1024485718843

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024485718843

Search

Navigation