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Influence of protein/glycan interaction on site-specific glycan heterogeneity.
Journal article

Influence of protein/glycan interaction on site-specific glycan heterogeneity.

  • Losfeld ME Department of Biology, Institute of Microbiology, Swiss Federal Institute of Technology (ETH) Zürich, Zürich, Switzerland.
  • Scibona E Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology ETH Zürich, Zürich, Switzerland.
  • Lin CW Department of Biology, Institute of Microbiology, Swiss Federal Institute of Technology (ETH) Zürich, Zürich, Switzerland.
  • Villiger TK Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology ETH Zürich, Zürich, Switzerland.
  • Gauss R Department of Biology, Institute of Microbiology, Swiss Federal Institute of Technology (ETH) Zürich, Zürich, Switzerland.
  • Morbidelli M Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology ETH Zürich, Zürich, Switzerland.
  • Aebi M Department of Biology, Institute of Microbiology, Swiss Federal Institute of Technology (ETH) Zürich, Zürich, Switzerland; markus.aebi@micro.biol.ethz.ch.
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  • 2017-07-07
Published in:
  • FASEB journal : official publication of the Federation of American Societies for Experimental Biology. - 2017
Golgi processing
enzymatic flux
glycobiology
glycoengineering
glycoprotein maturation
Animals
CHO Cells
Cricetulus
Endoplasmic Reticulum
Glycoproteins
Glycosylation
Golgi Apparatus
Oligosaccharides
Polysaccharides
Recombinant Proteins
English To study how the interaction between N-linked glycans and the surrounding amino acids influences oligosaccharide processing, we used protein disulfide isomerase (PDI), a glycoprotein bearing 5 N-glycosylation sites, as a model system and expressed it transiently in a Chinese hamster ovary (CHO)-S cell line. PDI was produced as both secreted Sec-PDI and endoplasmic reticulum-retained glycoprotein (ER)-PDI, to study glycan processing by ER and Golgi resident enzymes. Quantitative site-specific glycosylation profiles were obtained, and flux analysis enabled modeling site-specific glycan processing. By altering the primary sequence of PDI, we changed the glycan/protein interaction and thus the site-specific glycoprofile because of the improved enzymatic fluxes at enzymatic bottlenecks. Our results highlight the importance of direct interactions between N-glycans and surface-exposed amino acids of glycoproteins on processing in the ER and the Golgi and the possibility of changing a site-specific N-glycan profile by modulating such interactions and thus the associated enzymatic fluxes. Altering the primary protein sequence can therefore be used to glycoengineer recombinant proteins.-Losfeld, M.-E., Scibona, E., Lin, C.-W., Villiger, T. K., Gauss, R., Morbidelli, M., Aebi, M. Influence of protein/glycan interaction on site-specific glycan heterogeneity.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/57605
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