Selective N-glycan editing on living cell surfaces to probe glycoconjugate function.

Feng Tang, Mang Zhou, Ken Qin, Wei Shi, Ansor Yashinov, Yang Yang, Liyun Yang, Dongliang Guan, Lei Zhao, Yubo Tang, Yujie Chang, Lifen Zhao, Huaiyu Yang, Hu Zhou, Ruimin Huang, Wei Huang

Nature chemical biology 2020 Jul

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Cell surfaces are glycosylated in various ways with high heterogeneity, which usually leads to ambiguous conclusions about glycan-involved biological functions. Here, we describe a two-step chemoenzymatic approach for N-glycan-subtype-selective editing on the surface of living cells that consists of a first 'delete' step to remove heterogeneous N-glycoforms of a certain subclass and a second 'insert' step to assemble a well-defined N-glycan back onto the pretreated glyco-sites. Such glyco-edited cells, carrying more homogeneous oligosaccharide structures, could enable precise understanding of carbohydrate-mediated functions. In particular, N-glycan-subtype-selective remodeling and imaging with different monosaccharide motifs at the non-reducing end were successfully achieved. Using a combination of the expression system of the Lec4 CHO cell line and this two-step glycan-editing approach, opioid receptor delta 1 (OPRD1) was investigated to correlate its glycostructures with the biological functions of receptor dimerization, agonist-induced signaling and internalization.

Citation

Feng Tang, Mang Zhou, Ken Qin, Wei Shi, Ansor Yashinov, Yang Yang, Liyun Yang, Dongliang Guan, Lei Zhao, Yubo Tang, Yujie Chang, Lifen Zhao, Huaiyu Yang, Hu Zhou, Ruimin Huang, Wei Huang. Selective N-glycan editing on living cell surfaces to probe glycoconjugate function. Nature chemical biology. 2020 Jul;16(7):766-775