title: Liberation of Protein-Specific Glycosylation Information for Glycan Analysis by Exonuclease III-Aided Recycling Hybridization.
abstract: A strategy for information liberation of protein-specific glycosylation is designed via an exonuclease III-aided recycling "hybridization and cleavage" process with glycan and protein probes, which achieves homogeneous quantification of cell surface glycan. The protein probe contains matching and spacer DNA sequences and an aptamer specific to target protein. The glycan probe contains a complementary sequence modified with neighboring fluorescein and quencher, a spacer sequence, and a dibenzocyclooctyne-amine end to bind azide-tagged glycan. Upon sequential binding to their targets, the complementary sequences of two probes approach enough for their hybridization, which leads to the cleavage of hybridized glycan probe by exonuclease III and followed recycling "hybridization and cleavage" process of protein probe with other adjacent glycan probes to release the labeled fluorescein for obtaining the information on protein-specific glycosylation. This protocol has been used to in situ quantify EpCAM-specific sialic acid on MCF-7 cell surface and monitor its variation during drug treatment. This work demonstrates a powerful quantification tool for research of glycosylation.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing 210023, People's Republic of China.
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing 210023, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Ding
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing 210023, People's Republic of China.
authors.forename: Wanyao
authors.initials: W
authors.lastname: Song
authors.affiliation: Department of Pharmaceutical and Biological Chemistry, UCL School of Pharmacy, University College London , London WC1N 1AX, U.K.
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing 210023, People's Republic of China.
authors.forename: Huangxian
authors.initials: H
authors.lastname: Ju
mesh_terms: D005090:Exodeoxyribonucleases / Q000378:metabolism*; D006031:Glycosylation; D011134:Polysaccharides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
chemical_list: D011134:Polysaccharides; D011506:Proteins; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III