Sequence-Specific Mucins for Glycocalyx Engineering.

Area of Science:

• Biochemistry
• Molecular Biology
• Glycobiology

Background:

• Developing customized mucin glycoproteins for precise control over the glycocalyx in eukaryotic cells remains challenging.
• Existing methods lack the stability and controllability required for synthetic mucin engineering.

Purpose of the Study:

• To establish a versatile platform for the stable and controllable synthesis of customized mucin glycoproteins.
• To create a library of synthetic mucins with diverse properties for glycocalyx editing.

Main Methods:

• Utilized custom gene synthesis and a biology-by-parts approach for cDNA construction.
• Developed a library of swappable DNA bricks encoding mucin components (leader tags, anchors, motifs, reporters, repeats, domains).
• Constructed over 50 unique synthetic mucins with varied chemical, structural, and optical characteristics.

Main Results:

• Demonstrated sequence-specific effects on mucin glycosylation, showing peptide backbone influence on Tn-antigen extension to Core 1 and Core 2 structures.
• Identified that mucin cytoplasmic tail recycling motifs can modulate glycosylation.
• Established a modular system for readily constructing additional mucin permutations.

Conclusions:

• The developed mucin parts inventory provides a powerful tool for glycocalyx research.
• This platform facilitates the engineering of mucins with tailored properties for biotechnological applications.
• The findings offer new avenues for understanding and manipulating cellular glycosylation processes.