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Type: Journal article
Title: Isolation and characterisation of a recombinant precursor form of lysosomal acid α-glucosidase
Other Titles: Isolation and characterisation of a recombinant precursor form of lysosomal acid alpha-glucosidase
Author: Fuller, M.
Van der Ploeg, A.
Reuser, A.
Anson, D.
Hopwood, J.
Citation: Biochemical Journal, 1995; 234(3):903–909-
Publisher: Biochemical Society
Issue Date: 1995
ISSN: 0264-6021
Abstract: Glycogenosis type II (GSD II, Pompe disease) is an autosomal recessive lysosomal storage disease that results from a deficiency of acid alpha-glucosidase (GAA). Patients with this disorder are unable to break down lysosomal glycogen, which consequently accumulates in the lysosome. To evaluate enzyme replacement therapy for GSD II patients, we have expressed human GAA cDNA in Chinese hamster ovary-K1 cells utilising a vector that places the cDNA under the transcriptional control of the human polypeptide chain elongation factor 1 alpha gene promoter. A clonal cell line that secreted precursor recombinant GAA at approximately 18 was identified. The precursor recombinant GAA was purified to homogeneity, had a molecular mass of 110 kDa as measured by SDS/PAGE, and was shown to have pH optima and kinetic parameters similar to those of GAA purified from human tissues. The partial N-terminal amino acid sequence of recombinant GAA conformed to that derived from the nucleotide sequence of the cloned cDNA. The recombinant enzyme was taken up by cultured fibroblasts and skeletal muscle cells from GSD II patients, and was shown to correct the storage phenotype. Endocytosed GAA was localised to the lysosome and showed evidence of intracellular processing to a more mature form. Activity levels increased up to twice the normal value and uptake was prevented if cells were cultured in the presence of mannose 6-phosphate.
Keywords: Muscle, Skeletal
Cells, Cultured
CHO Cells
Glycogen Storage Disease Type II
Enzyme Precursors
Peptide Elongation Factor 1
Glucan 1,4-alpha-Glucosidase
Recombinant Proteins
Peptide Elongation Factors
Sequence Analysis
Protein Processing, Post-Translational
Amino Acid Sequence
Hydrogen-Ion Concentration
Molecular Sequence Data
Rights: @FEBS 1995
DOI: 10.1111/j.1432-1033.1995.903_a.x
Appears in Collections:Aurora harvest
Paediatrics publications

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