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Sci Adv. 2019 Nov 27;5(11):eaax8930. doi: 10.1126/sciadv.aax8930. eCollection 2019 Nov.

Mechanistic reconstruction of glycoprotein secretion through monitoring of intracellular N-glycan processing.

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Institute of Microbiology, Department of Biology, ETH Zürich, 8093 Zürich, Switzerland.
Institute for Chemical and Bioengineering, Department of Chemistry, ETH Zürich, 8093 Zürich, Switzerland.
Merck Healthcare, Biotech Process Sciences, Route de Fenil 25, 1804 Corsier-sur-Vevey, Switzerland.


N-linked glycosylation plays a fundamental role in determining the thermodynamic stability of proteins and is involved in multiple key biological processes. The mechanistic understanding of the intracellular machinery responsible for the stepwise biosynthesis of N-glycans is still incomplete due to limited understanding of in vivo kinetics of N-glycan processing along the secretory pathway. We present a glycoproteomics approach to monitor the processing of site-specific N-glycans in CHO cells. On the basis of a model-based analysis of structure-specific turnover rates, we provide a kinetic description of intracellular N-glycan processing along the entire secretory pathway. This approach refines and further extends the current knowledge on N-glycans biosynthesis and provides a basis to quantify alterations in the glycoprotein processing machinery.

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