High-Sensitivity and Low-Toxicity Fucose Probe for Glycan Imaging and Biomarker Discovery

Area of Science:

• Glycobiology
• Chemical Biology
• Biochemistry

Background:

• Fucose is a terminal sugar in glycoconjugates, regulating critical physiological and pathological processes like cancer and inflammation.
• Current methods for labeling and detecting fucose are inefficient, limiting research and clinical applications.
• There is a need for novel probes to accurately study fucose's role in biological systems.

Purpose of the Study:

• To synthesize and evaluate novel alkynyl-fucose analogs as efficient probes for fucose labeling and detection.
• To determine the substrate specificity of the synthesized analogs against fucosyltransferases.
• To assess the utility of the best-performing analog in cellular labeling and its potential for biomarker discovery.

Main Methods:

• Chemical synthesis of a novel series of alkynyl-fucose analogs.
• Evaluation of labeling efficiency and cytotoxicity of the synthesized analogs.
• Assay of analog substrate specificity against five fucosyltransferases.
• Confirmation of analog conversion to its GDP-derivative in cells.
• Mass spectrometry analysis for detection of labeled fucose in N-glycan core.
• Assessment of analog labeling in core-fucose-deficient mouse embryonic fibroblasts.

Main Results:

• 7-Alkynyl-fucose demonstrated the highest labeling efficiency and low cytotoxicity among the synthesized analogs.
• 7-Alkynyl-fucose served as the optimal substrate for all five tested fucosyltransferases.
• Cellular glycoproteins were labeled more efficiently with 7-alkynyl-fucose compared to existing probes.
• Mass spectrometry confirmed 7-alkynyl-fucose incorporation into the N-glycan core.
• The analog primarily labeled core fucose in mouse embryonic fibroblasts, as evidenced by reduced labeling in deficient cells.

Conclusions:

• 7-Alkynyl-fucose is a highly sensitive and effective probe for labeling and detecting fucose.
• This novel analog offers significant advantages over existing probes for glycoprotein analysis.
• 7-Alkynyl-fucose holds promise as a valuable tool for fundamental glycobiology research.
• The probe has potential applications in clinical settings for biomarker discovery.