1. Acid α-glucosidase was purified 3500-fold from rabbit muscle. 2. The enzyme was activated by cations, the degree of activation varying with the substrate. Enzyme action on glycogen was most strongly activated and activation was apparently of a non-competitive type. With rabbit liver glycogen as substrate, the relative Vmax. increased 15-fold, accompanied by an increase in Km from 8.3 to 68.6mm-chain end over the cation range 2–200mm-Na+ at pH4.5. Action on maltose was only moderately activated (1.3-fold, non-competitively) and action on maltotriose was marginally and competitively inhibited. 3. The pH optimum at 2mm-Na+ was 4.5 (maltose) and 5.1 (glycogen). Cation activation of enzyme action on glycogen was markedly pH-dependent. At 200mm-Na+, the pH optimum was 4.8 and activity was maximally stimulated in the range pH4.5–3.3. 4. Glucosidase action on maltosaccharides was associated with pronounced substrate inhibition at concentrations exceeding 5mm. Of the maltosaccharides tested, the enzyme showed a preference for p-nitrophenyl α-maltoside (Km 1.2mm) and maltotriose (Km 1.8mm). The extrapolated Km for enzyme action on maltose was 3.7mm. 5. The macromolecular polysaccharide substrate glycogen differed from linear maltosaccharide substrates in the kinetics of its interaction with the enzyme. Activity was markedly dependent on pH, cation concentration and polysaccharide structure. There was no substrate inhibition. 6. The enzyme exhibited constitutive α-1,6-glucanohydrolase activity. The Km for panose was 20mm. 7. The enzyme catalysed the total conversion of glycogen into glucose. The hydrolysis of α-1,6-linkages was apparently rate-limiting during the hydrolysis of glycogen. 8. Enzyme action on glycogen and maltose released the α-anomer of d-glucose. 9. The results are discussed in terms of the physiological role of acid α-glucosidase in lysosomal glycogen catabolism.
Research Article|October 01 1971
The substrate specificity of acid α-glucosidase from rabbit muscle
Biochem J (1971) 124 (4): 701-711.
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T. N. Palmer; The substrate specificity of acid α-glucosidase from rabbit muscle. Biochem J 1 October 1971; 124 (4): 701–711. doi: https://doi.org/10.1042/bj1240701
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