FEBS Letters | |
Properties of carbohydrate‐free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP‐glucose pyrophosphorylase activity | |
Lomako, Wieslawa M.1  Alonso, Miriam D.1  Lomako, Joseph1  Preiss, Jack2  Whelan, William J.1  | |
[1] Department of Biochemistry and Molecular Biology, University of Miami School of Medicine (M823), PO Box 016129, Miami, FL 33101, USA;Department of Biochemistry, Michigan State University, East Lansing, MI 48824, USA | |
关键词: Glycogenin; Apo-glycogenin; Carbohydrate-free glycogenin; | |
DOI : 10.1016/0014-5793(94)00962-7 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
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.
【 授权许可】
Unknown
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