Endogenous glycosyltransferase glucosylate lipids in flagella of Euglena

Shu-jen Chen, G. B. Bouck

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Flagella, intact deflagellated cells and isolated cell surfaces of the unicell, Euglena were separately assayed for glycosyltransferase activity by incubating these fractions with uridine diphosphate-[3H]glucose and isolating radiolabeled products. Most of the label was incorporated into lipophilic products, soluble in chloroform/methanol, which could be separated via thin layer chromatography or LH-60 chromatography into four distinct classes. The most polar of these products was extracted from flagella and purified by column chromatography for use as an in vitro substrate to identify flagella-associated glycosyltransferases. After flagella were treated with the detergent CHAPS, a soluble fraction was removed that was capable of glycosylation in solution. The glycosyltransferase(s) responsible for this activity were further enriched on sucrose or fructose gradients and ultimately identified on acrylamide gels through the combined use of nondenaturing gels, dial-[3H]uridine diphosphate binding, and fluorography. The enzyme had an apparent monomer molecular weight of 32,000 and consisted of four or fewer subunits. The occurrence of endogenous glycosyltransferase(s) in flagella suggests that modifications and/or assembly of the flagella surface can take place in situ in this organism.

Original languageEnglish (US)
Pages (from-to)1825-1835
Number of pages11
JournalJournal of Cell Biology
Volume98
Issue number5
DOIs
StatePublished - Jan 1 1984

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Euglena
Glycosyltransferases
Flagella
Lipids
Chromatography
Photofluorography
Gels
Uridine Diphosphate Glucose
Uridine Diphosphate
Acrylamide
Thin Layer Chromatography
Chloroform
Fructose
Glycosylation
Detergents
Methanol
Sucrose
Molecular Weight
Enzymes

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

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abstract = "Flagella, intact deflagellated cells and isolated cell surfaces of the unicell, Euglena were separately assayed for glycosyltransferase activity by incubating these fractions with uridine diphosphate-[3H]glucose and isolating radiolabeled products. Most of the label was incorporated into lipophilic products, soluble in chloroform/methanol, which could be separated via thin layer chromatography or LH-60 chromatography into four distinct classes. The most polar of these products was extracted from flagella and purified by column chromatography for use as an in vitro substrate to identify flagella-associated glycosyltransferases. After flagella were treated with the detergent CHAPS, a soluble fraction was removed that was capable of glycosylation in solution. The glycosyltransferase(s) responsible for this activity were further enriched on sucrose or fructose gradients and ultimately identified on acrylamide gels through the combined use of nondenaturing gels, dial-[3H]uridine diphosphate binding, and fluorography. The enzyme had an apparent monomer molecular weight of 32,000 and consisted of four or fewer subunits. The occurrence of endogenous glycosyltransferase(s) in flagella suggests that modifications and/or assembly of the flagella surface can take place in situ in this organism.",
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Endogenous glycosyltransferase glucosylate lipids in flagella of Euglena. / Chen, Shu-jen; Bouck, G. B.

In: Journal of Cell Biology, Vol. 98, No. 5, 01.01.1984, p. 1825-1835.

Research output: Contribution to journalArticle

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