Exploring Glycan-Lectin Interactions in Natural-Like Environments: A View Using NMR Experiments Inside Cell and on...

Area of Science:

• Biochemistry
• Molecular Biology
• Biophysics

Background:

• Glycan-mediated molecular recognition is vital for cellular processes.
• Nuclear Magnetic Resonance (NMR) is a key technique for studying glycan-lectin interactions in vitro.
• Existing NMR methods often use isolated components, not mimicking native cellular conditions.

Purpose of the Study:

• To develop and apply NMR methodologies for monitoring glycan-lectin interactions within cellular environments.
• To investigate glycan recognition from both receptor and ligand perspectives under native-like conditions.
• To establish proof-of-concept for NMR analysis of glycan recognition in situ.

Main Methods:

• Exploration of diverse NMR techniques, including 1H-15N HMQC NMR for intracellular observations and ligand-based STD-NMR for cell surface interactions.
• Utilized Danio Rerio oocytes for in-cell NMR experiments with galectin-7 and a glycomimetic ligand (TDG).
• Applied STD-NMR to study interactions of natural glycans and glycomimetics with cell surface-expressed Siglec-10.

Main Results:

• Successfully observed the 1H-15N HMQC NMR spectrum of folded galectin-7 inside Danio Rerio oocytes, a first for in-cell NMR of galectins.
• Demonstrated the use of a glycomimetic ligand (TDG) to facilitate intracellular galectin-7 NMR studies.
• Successfully applied STD-NMR to characterize glycan-Siglec-10 interactions on the cell surface.

Conclusions:

• NMR methodologies can be adapted to study glycan-lectin interactions in native-like cellular environments, both intracellularly and on the cell surface.
• These advanced NMR approaches provide new tools for understanding the biological roles of glycan recognition in situ.
• The study opens avenues for future NMR investigations into complex biological recognition events within their native cellular context.