PMID:7925977
| Citation |
Alonso, MD, Lomako, J, Lomako, WM, Whelan, WJ and Preiss, J (1994) Properties of carbohydrate-free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP-glucose pyrophosphorylase activity. FEBS Lett. 352:222-6 |
|---|---|
| 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 UDPxylose, 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. |
| Links | |
| Keywords |
Escherichia coli/enzymology; Escherichia coli/genetics; Gene Expression; Glucose/analysis; Glucosyltransferases; Glycogen/biosynthesis; Glycoproteins/biosynthesis; Glycoproteins/chemistry; Glycoproteins/genetics; Glycoproteins/metabolism; Glycosylation; Mutation/physiology; Recombinant Proteins/biosynthesis; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; UTP-Glucose-1-Phosphate Uridylyltransferase/metabolism; Uridine Diphosphate Glucose/metabolism; Uridine Diphosphate Xylose/metabolism |
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