A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes
- Author
- Petra Tiels (UGent) , Ekaterina Baranova, Kathleen Piens, Charlotte De Visscher (UGent) , Gwenda Pynaert, Wim Nerinckx (UGent) , Jan Stout, Franck Fudalej, Paco Hulpiau (UGent) , Simon Tännler, Steven Geysens, Annelies Van Hecke (UGent) , Albena Valevska, Wouter Vervecken, Han Remaut and Nico Callewaert (UGent)
- Organization
- 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
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-3087415
- MLA
- Tiels, Petra, et al. “A Bacterial Glycosidase Enables Mannose-6-Phosphate Modification and Improved Cellular Uptake of Yeast-Produced Recombinant Human Lysosomal Enzymes.” NATURE BIOTECHNOLOGY, vol. 30, no. 12, 2012, pp. 1225–31, doi:10.1038/nbt.2427.
- APA
- Tiels, P., Baranova, E., Piens, K., De Visscher, C., Pynaert, G., Nerinckx, W., … Callewaert, N. (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. https://doi.org/10.1038/nbt.2427
- Chicago author-date
- 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): 1225–31. https://doi.org/10.1038/nbt.2427.
- Chicago author-date (all authors)
- Tiels, Petra, Ekaterina Baranova, Kathleen Piens, Charlotte De Visscher, Gwenda Pynaert, Wim Nerinckx, Jan Stout, Franck Fudalej, Paco Hulpiau, Simon Tännler, Steven Geysens, Annelies Van Hecke, Albena Valevska, Wouter Vervecken, Han Remaut, and Nico Callewaert. 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. doi:10.1038/nbt.2427.
- 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.
- IEEE
- [1]P. Tiels et al., “A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes,” NATURE BIOTECHNOLOGY, vol. 30, no. 12, pp. 1225–1231, 2012.
@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 Tännler, 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}},
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}},
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://doi.org/10.1038/nbt.2427}},
volume = {{30}},
year = {{2012}},
}
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