[2-3H]Mannose-labeling and Analysis of N-linked Oligosaccharides

Marina Shenkman; Navit Ogen-Shtern; Gerardo Z Lederkremer

doi:10.21769/BioProtoc.2393

[2-³H]Mannose-labeling and Analysis of N-linked Oligosaccharides

Bio Protoc. 2017 Jul 20;7(14):e2393. doi: 10.21769/BioProtoc.2393.

Authors

Marina Shenkman¹, Navit Ogen-Shtern¹, Gerardo Z Lederkremer¹

Affiliation

¹ Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.

Abstract

Modifications of N-linked oligosaccharides of glycoproteins soon after their biosynthesis correlate to glycoprotein folding status. These alterations can be detected in a sensitive way by pulse-chase analysis of [2-³H]mannose-labeled glycoproteins, with enzymatic removal of labeled N-glycans, separation according to size by HPLC and radioactive detection in a scintillation counter.

Keywords: Endoplasmic reticulum associated degradation; Glycosylation; Mannose-labeling; Mannosidase; N-linked oligosaccharide.