Selective bioorthogonal probe for N-glycan hybrid structures.

Area of Science:

• Biochemistry
• Chemical Biology
• Glycobiology

Background:

• Metabolic labeling with tagged monosaccharides allows for glycoprotein and glycolipid tagging.
• Achieving selectivity in labeling is challenging due to shared monosaccharide precursor pathways.
• N-linked glycosylation, a complex posttranslational modification, yields three main N-glycan structures: oligomannose, hybrid, and complex.

Purpose of the Study:

• To develop a next-generation metabolic chemical reporter for selective labeling of hybrid N-glycan structures.
• To establish a general strategy for defining the selectivity of chemically tagged monosaccharide labeling.
• To utilize the reporter for imaging and fractionation to understand the localization and trafficking of hybrid N-glycan-bearing proteins.

Main Methods:

• Synthesis of the novel chemical reporter 1,3-Pr2-6-OTs GlcNAlk (MM-JH-1).
• Development of a general strategy to assess the selectivity of metabolic labeling.
• Application of MM-JH-1 for selective labeling and subsequent cellular imaging and fractionation.

Main Results:

• MM-JH-1 was successfully synthesized as a chemical reporter.
• The study established that MM-JH-1 is selectively incorporated into hybrid N-glycans.
• Imaging and fractionation experiments revealed the intracellular localization and trafficking of target proteins with hybrid N-glycans.

Conclusions:

• MM-JH-1 serves as a valuable tool for the selective metabolic labeling of hybrid N-glycans.
• This reporter facilitates the study of hybrid N-glycan-bearing proteins' cellular dynamics.
• The developed strategy provides a framework for designing selective metabolic labeling probes.