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A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes

(2012) NATURE BIOTECHNOLOGY. 30(12). p.1225-1231
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Abstract
Lysosomal storage diseases are treated with human lysosomal enzymes produced in mammalian cells. Such enzyme therapeutics contain relatively low levels of mannose-6-phosphate, which is required to target them to the lysosomes of patient cells. Here we describe a method for increasing mannose-6-phosphate modification of lysosomal enzymes produced in yeast. We identified a glycosidase from C. cellulans that 'uncaps' N-glycans modified by yeast-type mannose-Pi-6-mannose to generate mammalian-type N-glycans with a mannose-6-phosphate substitution. Determination of the crystal structure of this glycosidase provided insight into its substrate specificity. We used this uncapping enzyme together with alpha-mannosidase to produce in yeast a form of the Pompe disease enzyme alpha-glucosidase rich in mannose-6-phosphate. Compared with the currently used therapeutic version, this form of alpha-glucosidase was more efficiently taken up by fibroblasts from Pompe disease patients, and it more effectively reduced cardiac muscular glycogen storage in a mouse model of the disease.
Keywords
YARROWIA-LIPOLYTICA, STORAGE DISORDERS, N-LINKED OLIGOSACCHARIDES, MANNOSE 6-PHOSPHATE RECEPTORS, ACID ALPHA-GLUCOSIDASE, MOUSE MODEL, PICHIA-PASTORIS, POMPE-DISEASE, FABRY-DISEASE, SACCHAROMYCES-CEREVISIAE

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Citation

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Chicago
Tiels, Petra, Ekaterina Baranova, Kathleen Piens, Charlotte De Visscher, Gwenda Pynaert, Wim Nerinckx, Jan Stout, et al. 2012. “A Bacterial Glycosidase Enables Mannose-6-phosphate Modification and Improved Cellular Uptake of Yeast-produced Recombinant Human Lysosomal Enzymes.” Nature Biotechnology 30 (12): 1225–1231.
APA
Tiels, P., Baranova, E., Piens, K., De Visscher, C., Pynaert, G., Nerinckx, W., Stout, J., et al. (2012). A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes. NATURE BIOTECHNOLOGY, 30(12), 1225–1231.
Vancouver
1.
Tiels P, Baranova E, Piens K, De Visscher C, Pynaert G, Nerinckx W, et al. A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes. NATURE BIOTECHNOLOGY. 2012;30(12):1225–31.
MLA
Tiels, Petra, Ekaterina Baranova, Kathleen Piens, et al. “A Bacterial Glycosidase Enables Mannose-6-phosphate Modification and Improved Cellular Uptake of Yeast-produced Recombinant Human Lysosomal Enzymes.” NATURE BIOTECHNOLOGY 30.12 (2012): 1225–1231. Print.
@article{3087415,
  abstract     = {Lysosomal storage diseases are treated with human lysosomal enzymes produced in mammalian cells. Such enzyme therapeutics contain relatively low levels of mannose-6-phosphate, which is required to target them to the lysosomes of patient cells. Here we describe a method for increasing mannose-6-phosphate modification of lysosomal enzymes produced in yeast. We identified a glycosidase from C. cellulans that 'uncaps' N-glycans modified by yeast-type mannose-Pi-6-mannose to generate mammalian-type N-glycans with a mannose-6-phosphate substitution. Determination of the crystal structure of this glycosidase provided insight into its substrate specificity. We used this uncapping enzyme together with alpha-mannosidase to produce in yeast a form of the Pompe disease enzyme alpha-glucosidase rich in mannose-6-phosphate. Compared with the currently used therapeutic version, this form of alpha-glucosidase was more efficiently taken up by fibroblasts from Pompe disease patients, and it more effectively reduced cardiac muscular glycogen storage in a mouse model of the disease.},
  author       = {Tiels, Petra and Baranova, Ekaterina and Piens, Kathleen and De Visscher, Charlotte and Pynaert, Gwenda and Nerinckx, Wim and Stout, Jan and Fudalej, Franck and Hulpiau, Paco and T{\"a}nnler, Simon and Geysens, Steven and Van Hecke, Annelies and Valevska, Albena and Vervecken, Wouter and Remaut, Han and Callewaert, Nico},
  issn         = {1087-0156},
  journal      = {NATURE BIOTECHNOLOGY},
  language     = {eng},
  number       = {12},
  pages        = {1225--1231},
  title        = {A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes},
  url          = {http://dx.doi.org/10.1038/nbt.2427},
  volume       = {30},
  year         = {2012},
}

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