Stabilising normal and mis-sense variant alpha-glucosidase

FEBS Lett. 2006 Aug 7;580(18):4365-70. doi: 10.1016/j.febslet.2006.06.096. Epub 2006 Jul 10.

Abstract

alpha-Glucosidase (EC 3.2.1.3) is a lysosomal enzyme that hydrolyses alpha-1,4- and alpha-1,6-linkages of glycogen to produce free glucose. A deficiency in alpha-glucosidase activity results in glycogen storage disorder type II (GSD II), also called Pompe disease. Here, d-glucose was shown to be a competitive inhibitor of alpha-glucosidase and when added to culture medium at 6.0 g/L increased the production of this protein by CHO-K1 expression cells and stabilised the enzyme activity. D-Glucose also prevented alpha-glucosidase aggregation/precipitation and increased protein yield in a modified purification scheme. In fibroblast cells, from adult-onset GSD II patients, D-glucose increased the residual level of alpha-glucosidase activity, suggesting that a structural analogue of d-glucose may be used for enzyme enhancement therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Butyric Acid / pharmacology
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Enzyme Stability
  • Fibroblasts / enzymology
  • Glucose / pharmacology
  • Glycogen Storage Disease Type II / enzymology*
  • Glycogen Storage Disease Type II / genetics
  • Iduronidase / metabolism
  • Kinetics
  • Mutation, Missense
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Sulfatases / metabolism
  • alpha-Glucosidases / biosynthesis*
  • alpha-Glucosidases / genetics*
  • alpha-Glucosidases / metabolism

Substances

  • Recombinant Proteins
  • Butyric Acid
  • Sulfatases
  • alpha-Glucosidases
  • Iduronidase
  • Glucose