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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.

Author information

1
Institute of Microbiology, Department of Biology, ETH Zürich, 8093 Zürich, Switzerland.
2
Institute for Chemical and Bioengineering, Department of Chemistry, ETH Zürich, 8093 Zürich, Switzerland.
3
Merck Healthcare, Biotech Process Sciences, Route de Fenil 25, 1804 Corsier-sur-Vevey, Switzerland.

Abstract

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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