Lectin-specific targeting of lysosomal enzymes to reticuloendothelial cells.
The principles and methods used for enzymatic modification of the carbohydrate portion of glucocerebrosidase are similar to those performed by Ashwell and Morell, Stahl, and others. It is difficult to explain the lack of uptake of native enzyme through binding of the high-mannose type glycopeptide to Man/GlcNAc receptors since approximately 20% of the total oligosaccharides on the native enzyme are high mannose type. Possibly a requirement for multiple sites of attachment to the receptor is not met by a single high-mannose type oligosaccharide per molecule. Alternatively, the presence of complex type oligosaccharides on this enzyme, demonstrated by structural studies, may mask the mannose site and thus account for the poor uptake of native enzyme. The ability to successfully deglycosylate any protein or enzyme in order to specifically target a selected cell type requires that there be (1) an available source of pure enzyme; (2) specific exoglycosidases of high specific activity available either commercially or relatively easily purified; (3) chemical or biochemical means available for the testing of the product, preferably at each step; and (4) a means of separating the glycosidases used from the desired enzyme product. The characteristic and unique accumulation of glucocerebroside only in cells of the monocyte- histiocyte series, makes Gaucher's disease an excellent prototype for the study of enzyme replacement therapy. The principles demonstrated for the enzymatic deglycosylation of glucocerebrosidase may be applied to the cell-specific delivery of other glycoproteins as well. Other lysosomal diseases in which storage occurs in multiple cell types may be ameliorated by administration of macrophage-directed enzymes if, by so doing, storage material can be digested during the normal phagocytic turnover of senescent cells. Consideration of the kinetics of degradation and the structural features affecting the stability of enzymes in vivo are prerequisites to improving the bioengineering of targeted lysosomal enzymes. Animal and culture models have been developed for the study of glucocerebrosidase delivery to specific cell types and substrate degradation. Other studies have progressed toward a definition not only of the receptors at the plasma membrane involved in the internalization of exogenous enzyme, but also of internal receptors or properties of the lysosome responsible for intracellular protein trafficking. A complete understanding of the forces acting to direct endogenous or exogenously supplied enzyme to a given subcellular organelle will require a synthesis of experimental results from all areas of glycoprotein research.
Murray
G J
GJ
D000818:Animals; D001692:Biological Transport; D002241:Carbohydrates / Q000302:isolation & purification; D005260:Female; D005959:Glucosidases / Q000378:metabolism*; D005962:Glucosylceramidase / Q000302:isolation & purification / Q000378:metabolism*; D006026:Glycoside Hydrolases; D006801:Humans; D037102:Lectins; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D012157:Mononuclear Phagocyte System / Q000201:enzymology*; D009439:Neuraminidase; D010920:Placenta / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains
D002241:Carbohydrates; D037102:Lectins; D005959:Glucosidases; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D005962:Glucosylceramidase