Acid P-glucosidase

A-Butyl-l-deoxynojirimycin (131) HO VsY N Vv HO OH Acid P-glucosidase, human (GH 30) 2V3D 268... 

K. M. Osiecki-Newman, D. Fabbro, T. Dinur, S. Boas, S. Gatt, G. Legler, R. J. Desnick, and G. A. Grabowski, Human acid p-glucosidase Affinity purification of the normal placental and Gaucher disease splenic enzymes on N-alkyl-deoxynojirimycin-sepharose, Enzyme, 35 (1986) 147-153. 

B. Brumshtein, H. M. Greenblatt, T. D. Butters, Y. Shaaltiel, D. Aviezer, I. Sihnan, A. H. Futerman, and J. L. Sussman, Crystal structures of complexes of iV-butyl- and iV-nonyl-deoxynojirimycin bound to acid P-glucosidase. Insights into the mechanism of chemical chaperone action in Gaucher disease,... 

R. L. Lieberman, B. A. Wustman, P. Huertas, A. C. Powe, Jr., C. W. Pine, R. Khanna, M. G. Schlossmacher, D. Ringe, and G. A. Petsko, Structure of acid p-glucosidase with pharmacological chaperone provides insight into Gaucher disease, Nat. Chem. Biol., 3 (2007) 101-107. 

Gaucher s disease is an inborn error of sphingolipid metabolism caused by a deficiency of the lysosomal enzyme, acid p-glucosidase (EC 3.2.1.45) which is responsible for cleaving glucosylceramide (GlcCer) into glucose and... 

Acid P-glucosidase is routinely assayed with the fluorogenic 4-methyl-umbelliferyl-P-D-glucoside. Standardization of enzyme preparations is, however, problematic since the enzyme is stimulated by acidic phospholipids (Ho and Light, 1973 Dale et al., 1976 Peters et al., 1977a) or detergents (Ho, 1973 Dale et al., 1976 Peters et al., 1976) so that the absolute activity depends strongly on the amount and nature of such additions. 

Assay. Acid p-glucosidase hydrolyzes both water-soluble and lipid substrates. The activity, and hence the activation of this enzyme by cofactors, can therefore be determined with either kind of substrate. Due to the rapidity and convenience of assays with fluorogenic substrates, most workers prefer to use the commercially available 4-methylumbelli-feryl-p-D-glucoside. In some cases it may, however, be desirable, or even necessary, to confirm the results with the natural substrate, glucosylceramide. Both types of assays will therefore be described here. 

The interaction between acid p-glucosidase and activating protein cofactors depends on the presence of acidic lipids (Ho and Light, 1973), so a certain amount of such lipids has to be added to the assay system. (The partially purified enzyme preparations used contain frequently some lipids, including acidic ones.) On the other hand, higher concentrations of acidic lipids may stimulate the enzyme to almost maximal activity (Berent and Radin, 1981a) and may render protein cofactors obsolete. 

If a more purified preparation is desired, the enzyme, like acid P-glucosidase, can be converted into a water-soluble form by extraction with organic solvents (Tanaka and Suzuki, 1976,1977). However, the purification protocols published so far (Radin, 1972b Wenger et al., 1975) rely mainly on extraction with detergents. 

Fig. 3. Radiation inactivation size of acid p-glucosidase in normal ( ) and Gaucher (O) spleens as determined at 22°C with NBD-glucosylceramide as substrate. (From Maret et al., 1983.)...

Ho, MW and O Brien, JS (1971) Gaucher s disease deficiency of acid P-glucosidase and reconstitution of enzyme activity in vitro. Proc Natl Acad Sci USA, 68, 2810-2813. 

Oleuropein Hydroj lyrosol elenoic acid p-Glucosidase esterase Lact. plantamm... 

Different types of gel materials, such as polysaccharides, proteins and synthetic polymers, are now used to entrap biocatalysts. Among them, photo-crosslinkable resin prepolymer ENTP-4000 as shown in Eig. 7 is more useful compared to others. Entrapment of biocatalysts should be carried out under the illumination of near ultraviolet hght within 3-5 min, by which high temperatures, shifts of pH to extremely alkahne or acidic sides are avoided. ENTP-4000, hydrophobic photo-crosslinkable resin prepolymer, is one of the most suitable prepolymers for entrapment of p-glucosidase. Molecular weight of its main chain is about 4000. 

RL, Wustman BA, Huertas P, Powe AC, Pine CW, Khanna R, Schlossmacher MG Ringe D, Petsko GA. (2007) Stracture of acid beta-glucosidase with pharmacological chaperone provides insight into Gaucher disease. Nat Chem Biol 3 101-107. 

Amalfitano, A., Yie-Wylie, A. J., Hu, H., Dawson, T. L., Raben, N., Plotz, P. and Chen, Y. T. (1999). Systemic correction of the muscle disorder glycogen storage disease type II after hepatic targeting of a modified adenovirus vector encoding human acid-alpha-glucosidase. Proc. Natl. Acad. Sci. USA 96, 8861-8866. 

Bijvoet, A. G., Kroos, M. A., Pieper, F. R., Van, d., V, De Boer, H. A., Van der Ploeg, A. T., Verbeet, M. P. and Reuser, A. J. (1998). Recombinant human acid alpha-glucosidase High level production in mouse milk, biochemical characteristics, correction of enzyme deficiency in GSDII KO mice. Hum. Mol. Genet. 7, 1815-1824. 

Martin-Touaux, E., Puech, J. P., Chateau, D., Emiliani, C., Kremer, E. J., Raben, N., Tancini, B., Orlacchio, A., Kahn, A. and Poenaru, L. (2002). Muscle as a putative producer of acid alpha-glucosidase for glycogenosis type II gene therapy. Hum. Mol. Genet. 11, 1637-1645. 

Pauly, D. F., Fraites, T. J., Toma, C., Bayes, H. S., Huie, M. L., Hirschhorn, R., Plotz, P. H., Raben, N., Kessler, P. D. and Byrne, B. J. (2001). Intercellular transfer of the virally derived precursor form of acid alpha-glucosidase corrects the enzyme deficiency in inherited cardioskeletal myopathy Pompe disease. Hum. Gene Ther. 12(5), 527-538. 

Pauly, D. F., Johns, D. C., Matelis, L. A., Lawrence, J. H., Byrne, B. J. and Kessler, P. D. (1998). Complete correction of acid alpha-glucosidase deficiency in Pompe disease fibroblasts in vitro, and lysosomally targeted expression in neonatal rat cardiac and skeletal muscle. Gene Ther. 5, 473-480. 

Raben, N., Nagaraju, K., Lee, E., Kessler, P., Byrne, B., Lee, L., La Marca, M., King, C., Ward, J., Sauer, B. and Plotz, P. (1998). Targeted disruption of the acid alpha-glucosidase gene in mice causes an illness with critical features of both infantile and adult human glycogen storage disease type II. J. Biol. Chem. 273, 19086-19092. 

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