Mechanistic reconstruction of glycoprotein secretion through monitoring of intracellular N-glycan processing
Mechanistic reconstruction of glycoprotein secretion through monitoring of intracellular N-glycan processing
复制标题
DOI：
10.1126/sciadv.aax8930
复制
发表时间：
2019-11-01
期刊：
影响因子：
13.6
通讯作者：
Aebi, Markus
中科院分区：
文献类型：
--
作者：
Arigoni-Affolter, Ilaria;Scibona, Ernesto;Aebi, Markus
N-连接糖基化在决定蛋白质的热力学稳定性方面起着重要作用，并参与多种关键的生物学过程。由于对N-糖沿分泌途径的体内加工动力学的了解有限，对负责N-糖步生物合成的细胞内机制的机制了解仍然不完整。我们提出了一种糖蛋白组学方法来监测CHO细胞中位点特异性N-糖链的加工过程。在结构特异性周转率的模型分析的基础上，我们提供了沿着整个分泌途径的细胞内N-糖链加工的动力学描述。这种方法完善和进一步扩展了目前关于N-糖链生物合成的知识，并为量化糖蛋白加工机制中的变化提供了基础。
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.