Enzymatic characterization of human alpha1,3-fucosyltransferase Fuc-TVII synthesized in a B cell lymphoma cell line.
J Biol Chem
; 272(51): 31992-7, 1997 Dec 19.
Article
en En
| MEDLINE
| ID: mdl-9405391
The human alpha1,3-fucosyltransferase, Fuc-TVII, a key enzyme in the biosynthesis of selectin ligands, was expressed as a soluble protein-A chimeric form in a human B cell lymphoma cell line, Namalwa KJM-1, and purified using IgG-Sepharose. The enzymatic properties of recombinant soluble Fuc-TVII were then examined. Its enzyme activity was highest at pH 7.5, and the presence of 25 mM Mn2+ was required for full activity. Fuc-TVII exhibits an acceptor specificity restricted to alpha2,3-sialylated type 2 oligosaccharides, and the apparent Km values for alpha2,3-sialyl lacto-N-neotetraose and GDP-fucose were 3.08 mM and 16.4 microM, respectively. The inhibitory effects of various nucleotides on the activity of Fuc-TVII reflected its donor specificity for the nucleotide portion of GDP. Fuc-TVII was demonstrated to be useful for the synthesis of a sialyl Lewis x hexasaccharide from lacto-N-neotetraose in combination with an alpha2, 3-sialyltransferase, ST3Gal IV. Polyethylene glycols enhanced the thermal stability of Fuc-TVII, leading to increased formation of the reaction product.
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Banco de datos:
MEDLINE
Asunto principal:
Linfoma de Burkitt
/
Fucosiltransferasas
Límite:
Humans
Idioma:
En
Revista:
J Biol Chem
Año:
1997
Tipo del documento:
Article
País de afiliación:
Japón