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


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.

SUBMITTER: Arigoni-Affolter I 

PROVIDER: S-EPMC6881162 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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

Arigoni-Affolter Ilaria I   Scibona Ernesto E   Lin Chia-Wei CW   Brühlmann David D   Souquet Jonathan J   Broly Hervé H   Aebi Markus M  

Science advances 20191127 11


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. O  ...[more]

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