d-Galactosyltransferase and its endogenous substrates in chick embryo fibroblasts transformed by rous sarcoma virus

Daniel K. Podolsky, Deborah A. Fournier, Kurt J. Isselbacher

Research output: Contribution to journalArticle

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Abstract

UDP=d-galactose: 2-acetamido-2-deoxy-β-d-glucopyranosyl 4=β-d-galactosyltransferase (GalTase) activity was purified, from primary chick embryo fibroblast (CEF) transformed by a temperature-sensitive, Rous sarcoma virus mutant (CEF-RSV), by chromatography on an affinity resin prepared with monoclonal antibodies to GalTase. Cellular glycopeptides from CEF, as well as CEF-RSV, maintained at permissive (37°) [CEF-RSV (37°)] and nonpermissive temperatures (41°) [CEF-RSV (41°)], were solubilized and galactosylated in vitro by incubation with purified GalTase substrates, composed of at least six discrete complex glycopeptides having bi- to tetra-antennary structures. The glycopeptides isolated from transformed cells, CEF-RSV (37°), included the six types observed in nontransformed cells, but demonstrated alterations in their relative amounts, including an increase in the content of a glycopeptide containing 3 mannose and 4 glucosamine residues. Furthermore, two additional complex-type glycopeptides were isolated from CEF- but demonstrated alterations in their relative amounts, including an increase in the content of a glycopeptide containing 3 mannose and 4 glucosamine residues. Furthermore, two additional complex type glycopeptides were isolated from CEF-RSV (37°). These malignant transformation-related glycopeptides were partially characterized and found to represent tri- and tetra-antennary complex glycopeptides. Endogenous galactosylation appeared to have occurred in a branched, nonspecific manner in these transformed cell-derived glycopeptides. These findings indicate that transformed cells may contain a greater preponderance of more highly branched, complex oligosaccharides which are randomly galactosylated at non-reducing termini by cellular GalTase.

Original languageEnglish (US)
Pages (from-to)225-239
Number of pages15
JournalCarbohydrate Research
Volume149
Issue number1
DOIs
StatePublished - Jun 1 1986

Fingerprint

Galactosyltransferases
Rous sarcoma virus
Glycopeptides
Chick Embryo
Fibroblasts
Viruses
Substrates
Glucosamine
Mannose
Uridine Diphosphate Galactose
Temperature
Uridine Diphosphate
Chromatography
Oligosaccharides
Galactose
Affinity Chromatography
Resins
Monoclonal Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry

Cite this

d-Galactosyltransferase and its endogenous substrates in chick embryo fibroblasts transformed by rous sarcoma virus. / Podolsky, Daniel K.; Fournier, Deborah A.; Isselbacher, Kurt J.

In: Carbohydrate Research, Vol. 149, No. 1, 01.06.1986, p. 225-239.

Research output: Contribution to journalArticle

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