Check out our latest paper Optimizing phosphate couplings for dolichyl diphosphochitobiose to enable protein N-glycosylation studies in HELVETICA!

Abstract

Herein we report an optimized synthesis for dolichyl diphosphochitobiose (GlcNAc2-PP-Dol25), a probe useful for biochemical and structural studies ofprotein N-glycosylation in eukaryotic cells. We improved three phosphate coupling steps in terms of yields and reaction times, namely chitobiose phosphorylation, dolichol phosphorylation, and phosphate-phosphate coupling, by adjusting reagents and conditions. We also developed an efficient preparative reverse-phase HPLC purification protocol followed by ion exchange step to obtain the pure product as a stable sodium salt. These optimized procedures ensure a reliable supply of GlcNAc2-PP-Dol25 for enzymatic studies.

Author(s): Matheus A.Meirelles and Jean-Louis Reymond