Functional Purification and Characterization of a GDP-fucose: Beta-N-acetylglucosamine (Fuc to Asn Linked GlcNAc) Alpha 1,3-fucosyltransferase from Mung Beans

Overview

Journal Glycoconj J

Publisher Springer

Specialties Biochemistry
Endocrinology

Date 1995 Dec 1

PMID 8748155

Citations 12

Authors

E Staudacher

T Dalik

P Wawra

F Altmann

L Marz

Affiliations

Soon will be listed here.

Abstract

An alpha 1,3-fucosyltransferase was purified 3000-fold from mung bean seedlings by chromatography on DE 52 cellulose and Affigel Blue, by chromatofocusing, gelfiltration and affinity chromatography resulting in an apparently homogenous protein of about 65 kDa on SDS-PAGE. The enzyme transferred fucose from GDP-fucose to the Asn-linked N-acetylglucosaminyl residue of an N-glycan, forming an alpha 1,3-linkage. The enzyme acted upon N-glycopeptides and related oligosaccharides with the glycan structure GlcNAc2Man3 GlcNAc2. Fucose in alpha 1,6-linkage to the asparagine-linked GlcNAc had no effect on the activity. No transfer to N-glycans was observed when the terminal GlcNAc residues were either absent or substituted with galactose. N-acetyllactosamine, lacto-N-biose and N-acetylchito-oligosaccharides did not function as acceptors for the alpha 1,3-fucosyltransferase. The transferase exhibited maximal activity at pH 7.0 and a strict requirement for Mn2+ or Zn2+ ions. The enzyme's activity was moderately increased in the presence of Triton X-100. It was not affected by N-ethylmaleimide.

Citing Articles

Plant glyco-biotechnology on the way to synthetic biology.

Loos A, Steinkellner H Front Plant Sci. 2014; 5:523.

PMID: 25339965 PMC: 4189330. DOI: 10.3389/fpls.2014.00523.

A unique beta1,3-galactosyltransferase is indispensable for the biosynthesis of N-glycans containing Lewis a structures in Arabidopsis thaliana.

Strasser R, Bondili J, Vavra U, Schoberer J, Svoboda B, Glossl J Plant Cell. 2007; 19(7):2278-92.

PMID: 17630273 PMC: 1955701. DOI: 10.1105/tpc.107.052985.

An antibody produced in tobacco expressing a hybrid beta-1,4-galactosyltransferase is essentially devoid of plant carbohydrate epitopes.

Bakker H, Rouwendal G, Karnoup A, Florack D, Stoopen G, Helsper J Proc Natl Acad Sci U S A. 2006; 103(20):7577-82.

PMID: 16675551 PMC: 1472489. DOI: 10.1073/pnas.0600879103.

Arabidopsis thaliana beta1,2-xylosyltransferase: an unusual glycosyltransferase with the potential to act at multiple stages of the plant N-glycosylation pathway.

Bencur P, Steinkellner H, Svoboda B, Mucha J, Strasser R, Kolarich D Biochem J. 2005; 388(Pt 2):515-25.

PMID: 15686448 PMC: 1138959. DOI: 10.1042/BJ20042091.

A genetic and structural analysis of the N-glycosylation capabilities.

Leonard R, Kolarich D, Paschinger K, Altmann F, Wilson I Plant Mol Biol. 2004; 55(5):631-44.

PMID: 15604706 DOI: 10.1007/s11103-004-1558-3.

References

Staudacher E, Altmann F, Marz L, Hard K, Kamerling J, Vliegenthart J . Alpha 1-6(alpha 1-3)-difucosylation of the asparagine-bound N-acetylglucosamine in honeybee venom phospholipase A2. Glycoconj J. 1992; 9(2):82-5. DOI: 10.1007/BF00731703. View

Sasaki K, Kurata K, Funayama K, Nagata M, Watanabe E, Ohta S . Expression cloning of a novel alpha 1,3-fucosyltransferase that is involved in biosynthesis of the sialyl Lewis x carbohydrate determinants in leukocytes. J Biol Chem. 1994; 269(20):14730-7. View

Kurosaka A, Yano A, Itoh N, Kuroda Y, Nakagawa T, Kawasaki T . The structure of a neural specific carbohydrate epitope of horseradish peroxidase recognized by anti-horseradish peroxidase antiserum. J Biol Chem. 1991; 266(7):4168-72. View

Faye L, Gomord V, Chrispeels M . Affinity purification of antibodies specific for Asn-linked glycans containing alpha 1-->3 fucose or beta 1-->2 xylose. Anal Biochem. 1993; 209(1):104-8. DOI: 10.1006/abio.1993.1088. View

Tezuka K, Hayashi M, Ishihara H, Akazawa T, Takahashi N . Studies on synthetic pathway of xylose-containing N-linked oligosaccharides deduced from substrate specificities of the processing enzymes in sycamore cells (Acer pseudoplatanus L.). Eur J Biochem. 1992; 203(3):401-13. DOI: 10.1111/j.1432-1033.1992.tb16564.x. View

Szumilo T, Kaushal G, Elbein A . Demonstration of GlcNAc transferase I in plants. Biochem Biophys Res Commun. 1986; 134(3):1395-403. DOI: 10.1016/0006-291x(86)90404-3. View

Szumilo T, Kaushal G, Elbein A . Purification and properties of glucosidase I from mung bean seedlings. Arch Biochem Biophys. 1986; 247(2):261-71. DOI: 10.1016/0003-9861(86)90583-7. View

Natsuka S, Gersten K, Zenita K, Kannagi R, Lowe J . Molecular cloning of a cDNA encoding a novel human leukocyte alpha-1,3-fucosyltransferase capable of synthesizing the sialyl Lewis x determinant. J Biol Chem. 1994; 269(24):16789-94. View

Natunen J, Niemela R, Penttila L, Seppo A, Ruohtula T, Renkonen O . Enzymatic synthesis of two lacto-N-neohexaose-related Lewis x heptasaccharides and their separation by chromatography on immobilized wheat germ agglutinin. Glycobiology. 1994; 4(5):577-83. DOI: 10.1093/glycob/4.5.577. View

10.

Staudacher E, Altmann F, Glossl J, Marz L, SCHACHTER H, Kamerling J . GDP-fucose: beta-N-acetylglucosamine (Fuc to (Fuc alpha 1----6GlcNAc)-Asn-peptide)alpha 1----3-fucosyltransferase activity in honeybee (Apis mellifica) venom glands. The difucosylation of asparagine-bound N-acetylglucosamine. Eur J Biochem. 1991; 199(3):745-51. DOI: 10.1111/j.1432-1033.1991.tb16179.x. View

11.

Voynow J, Kaiser R, Scanlin T, GLICK M . Purification and characterization of GDP-L-fucose-N-acetyl beta-D-glucosaminide alpha 1----6fucosyltransferase from cultured human skin fibroblasts. Requirement of a specific biantennary oligosaccharide as substrate. J Biol Chem. 1991; 266(32):21572-7. View

12.

Szumilo T, Kaushal G, Elbein A . Purification and properties of the glycoprotein processing N-acetylglucosaminyltransferase II from plants. Biochemistry. 1987; 26(17):5498-505. DOI: 10.1021/bi00391a043. View

13.

Tretter V, Altmann F, KUBELKA V, Marz L, Becker W . Fucose alpha 1,3-linked to the core region of glycoprotein N-glycans creates an important epitope for IgE from honeybee venom allergic individuals. Int Arch Allergy Immunol. 1993; 102(3):259-66. DOI: 10.1159/000236534. View

14.

Szumilo T, Kaushal G, Hori H, Elbein A . Purification and Properties of a Glycoprotein Processing alpha-Mannosidase from Mung Bean Seedlings. Plant Physiol. 1986; 81(2):383-9. PMC: 1075345. DOI: 10.1104/pp.81.2.383. View

15.

Crawley S, Hindsgaul O, Ratcliffe R, Lamontagne L, Palcic M . A plant fucosyltransferase with human Lewis blood-group specificity. Carbohydr Res. 1989; 193:249-56. DOI: 10.1016/0008-6215(89)85123-7. View

16.

Johnson K, Chrispeels M . Substrate Specificities of N-Acetylglucosaminyl-, Fucosyl-, and Xylosyltransferases that Modify Glycoproteins in the Golgi Apparatus of Bean Cotyledons. Plant Physiol. 1987; 84(4):1301-8. PMC: 1056769. DOI: 10.1104/pp.84.4.1301. View

17.

Kaushal G, Szumilo T, Pastuszak I, Elbein A . Purification to homogeneity and properties of mannosidase II from mung bean seedlings. Biochemistry. 1990; 29(8):2168-76. DOI: 10.1021/bi00460a030. View

18.

Staudacher E, KUBELKA V, Marz L . Distinct N-glycan fucosylation potentials of three lepidopteran cell lines. Eur J Biochem. 1992; 207(3):987-93. DOI: 10.1111/j.1432-1033.1992.tb17134.x. View

19.

de Vries T, van den Eijnden D . Biosynthesis of sialyl-oligomeric-Lewisx and VIM-2 epitopes: site specificity of human milk fucosyltransferase. Biochemistry. 1994; 33(33):9937-44. DOI: 10.1021/bi00199a016. View

20.

KUBELKA V, Altmann F, Staudacher E, Tretter V, Marz L, Hard K . Primary structures of the N-linked carbohydrate chains from honeybee venom phospholipase A2. Eur J Biochem. 1993; 213(3):1193-204. DOI: 10.1111/j.1432-1033.1993.tb17870.x. View