Efficient Preparation and Bioactivity Evaluation of Glycan-Defined Glycoproteins

Owing to the generation of heterogeneous glycoproteins in cells, it is highly difficult to study glycoprotein-mediated biological events and to develop biomedical agents. Thus, general and efficient methods to prepare homogeneous glycoproteins are in high demand. Herein, we report a general method f...

Full description

Saved in:
Bibliographic Details
Published in:ACS chemical biology 2021-10, Vol.16 (10), p.1930-1940
Main Authors: Hyun, Ji Young, Kim, Sanggil, Lee, Chang-Hee, Lee, Hyun Soo, Shin, Injae
Format: Article
Language:English
Subjects:
Alkynes - chemistry
Azides - chemistry
beta-N-Acetylhexosaminidases - chemical synthesis
beta-N-Acetylhexosaminidases - genetics
beta-N-Acetylhexosaminidases - metabolism
Biocatalysis
Click Chemistry
Fibroblasts - metabolism
G(M2) Ganglioside - metabolism
Glycoproteins - chemical synthesis
Glycoproteins - genetics
Glycoproteins - metabolism
Glycosylation
Humans
Lectins - metabolism
Lysosomes - metabolism
Mutation
Polysaccharides - chemistry
Polysaccharides - genetics
Protein Processing, Post-Translational
THP-1 Cells
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Owing to the generation of heterogeneous glycoproteins in cells, it is highly difficult to study glycoprotein-mediated biological events and to develop biomedical agents. Thus, general and efficient methods to prepare homogeneous glycoproteins are in high demand. Herein, we report a general method for the efficient preparation of homogeneous glycoproteins that utilizes a combination of genetic code expansion and chemoselective ligation techniques. In the protocol to produce glycan-defined glycoproteins, an alkyne tag-containing protein, generated by genetic encoding of an alkynylated unnatural amino acid, was quantitatively coupled via click chemistry to versatile azide-appended glycans. The glycoproteins produced by the present strategy were found to recognize mammalian cell-surface lectins and enter the cells through lectin-mediated internalization. Also, cell studies exhibited that the glycoprotein containing multiple mannose-6-phosphate residues enters diseased cells lacking specific lysosomal glycosidases by binding to the cell-surface M6P receptor, and subsequently migrates to lysosomes for efficient degradation of stored glycosphingolipids.
ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.0c00629