Stabilising normal and mis-sense variant alpha-glucosidase.
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.
Kakavanos
Revecca
R
Department of Genetic Medicine, Lysosomal Diseases Research Unit, Children Youth and Women's Health Service, North Adelaide, SA 5006, Australia.
Hopwood
John J
JJ
Lang
Debbie
D
Meikle
Peter J
PJ
Brooks
Doug A
DA
D000818:Animals; D020148:Butyric Acid / Q000494:pharmacology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004795:Enzyme Stability; D005347:Fibroblasts / Q000201:enzymology; D005947:Glucose / Q000494:pharmacology; D006009:Glycogen Storage Disease Type II / Q000201:enzymology* / Q000235:genetics; D007068:Iduronidase / Q000378:metabolism; D007700:Kinetics; D020125:Mutation, Missense; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013429:Sulfatases / Q000378:metabolism; D000520:alpha-Glucosidases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D020148:Butyric Acid; D013429:Sulfatases; D000520:alpha-Glucosidases; D007068:Iduronidase; D005947:Glucose