Properties of carbohydrate-free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP-glucose pyrophosphorylase activity

Miriam D. Alonso, Joseph Lomako, Wieslawa M. Lomako, William J. Whelan, Jack Preiss

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Glycogenin, the self-glucosylating primer for glycogen synthesis is expressed in wild-type E. coli as a recombinant protein in an already partly glucosylated form owing to the presence of its substrate UDP-glucose. By using an E. coli mutant strain lacking in UDP-glucose pyrophosphorylase activity, we have succeeded in expressing carbohydrate-free glycogenin (apo-glycogenin) in good yield. When provided with UDPxy-lose, it autocatalytically adds 1 xylose residue. With UDP-glucose, an average of 8 glucose residues are added. However release of the self-synthesized maltosaccharide chains with isoamylase reveals them to be a mixture. Chains as long as 11 glucose residues (maltoundecaose) are present. The ability of recombinant apo-glycogenin to self-glucosylate is further proof that a separate enzyme is not needed for the addition of the first glucose residue to Tyr-194 of the protein.

Original languageEnglish
Pages (from-to)222-226
Number of pages5
JournalFEBS Letters
Volume352
Issue number2
DOIs
StatePublished - Sep 26 1994

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UTP-Glucose-1-Phosphate Uridylyltransferase
Escherichia coli
Carbohydrates
Uridine Diphosphate Glucose
Glucose
Isoamylase
Xylose
Glycogen
Recombinant Proteins
glycogenin
Substrates
Enzymes
Proteins

Keywords

  • Apo-glycogenin
  • Carbohydrate-free glycogenin
  • Glycogenin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Properties of carbohydrate-free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP-glucose pyrophosphorylase activity. / Alonso, Miriam D.; Lomako, Joseph; Lomako, Wieslawa M.; Whelan, William J.; Preiss, Jack.

In: FEBS Letters, Vol. 352, No. 2, 26.09.1994, p. 222-226.

Research output: Contribution to journalArticle

Alonso, Miriam D. ; Lomako, Joseph ; Lomako, Wieslawa M. ; Whelan, William J. ; Preiss, Jack. / Properties of carbohydrate-free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP-glucose pyrophosphorylase activity. In: FEBS Letters. 1994 ; Vol. 352, No. 2. pp. 222-226.
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