Identification of Glycosyltransferase 8 Family Members as Xylosyltransferases Acting on O-Glucosylated Notch Epidermal Growth Factor Repeats

Identification of Glycosyltransferase 8 Family Members as Xylosyltransferases Acting on O-Glucosylated Notch Epidermal Growth Factor Repeats Full article

Journal

Journal of Biological Chemistry
ISSN: 1083-351X , E-ISSN: 0021-9258

Output data

Year: 2010, Volume: 285, Number: 3, Pages: 1582-1586 Pages count : 5 DOI: 10.1074/jbc.c109.065409

Authors

Sethi Maya K. ¹ , Buettner Falk F.R. ^1,2 , Krylov Vadim B. ³ , Takeuchi Hideyuki ⁴ , Nifantiev Nikolay E. ³ , Haltiwanger Robert S. ⁴ , Gerardy-Schahn Rita ¹ , Bakker Hans ¹

Affiliations

1	Department of Cellular Chemistry, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany
2	Department of Infectious Diseases, Institute for Microbiology, University of Veterinary Medicine Hannover, 30173 Hannover, Germany
3	Laboratory of Glycoconjugate Chemistry, N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospect 47, 119991 Moscow, Russian Federation
4	Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, Stony Brook University, Stony Brook, New York 11794-5215

The epidermal growth factor repeats of the Notch receptor are extensively glycosylated with three different O-glycans. O-Fucosylation and elongation by the glycosyltransferase Fringe have been well studied and shown to be essential for proper Notch signaling. In contrast, biosynthesis of O-glucose and O-N-acetylglucosamine is less well understood. Recently, the isolation of the Drosophila mutant rumi has shown that absence of O-glucose impairs Notch function. O-Glucose is further extended by two contiguous α1,3-linked xylose residues. We have identified two enzymes of the human glycosyltransferase 8 family, now named GXYLT1 and GXYLT2 (glucoside xylosyltransferase), as UDP-d-xylose:β-d-glucoside α1,3-d-xylosyltransferases adding the first xylose. The enzymes are specific for β-glucose-terminating acceptors and UDP-xylose as donor substrate. Generation of the α1,3-linkage was confirmed by nuclear magnetic resonance. Activity on a natural acceptor could be shown by in vitro xylosylation of a Notch fragment expressed in a UDP-xylose-deficient cell line and in vivo by co-expression of the enzymes and the Notch fragment in insect cells followed by mass spectrometric analysis of peptide fragments.

Sethi M.K. , Buettner F.F.R. , Krylov V.B. , Takeuchi H. , Nifantiev N.E. , Haltiwanger R.S. , Gerardy-Schahn R. , Bakker H.
Identification of Glycosyltransferase 8 Family Members as Xylosyltransferases Acting on O-Glucosylated Notch Epidermal Growth Factor Repeats
Journal of Biological Chemistry. 2010. V.285. N3. P.1582-1586. DOI: 10.1074/jbc.c109.065409 WOS Scopus OpenAlex

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