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Positional Scanning MUC1 Glycopeptide Library Reveals the Importance of PDTR Epitope Glycosylation for Lectin Binding
YashoNandini Singh1, Maria C Rodriguez Benavente1, Mohammed H Al-Huniti1
1From the Department of Chemistry and Biochemistry, Charles E. Schmidt College of Science , Florida Atlantic University , 777 Glades Road , Boca Raton , Florida 33431 , United States.
Abstract:
One of the main barriers to explaining the functional significance of glycan-based changes in cancer is the natural epitope heterogeneity found on the surface of cancer cells. To help address this knowledge gap, we focused on designing synthetic tools to explore the role of tumor-associated glycans of MUC1 in the formation of metastasis via association with lectins. In this study, we have synthesized for the first time a MUC1-derived positional scanning synthetic glycopeptide combinatorial library (PS-SGCL) that vary in number and location of cancer-associated Tn antigen using the "tea bag" approach. The determination of the isokinetic ratios necessary for the equimolar incorporation of (glyco)amino acids mixtures to resin-bound amino acid was determined, along with developing an efficient protocol for on resin deprotection of O-acetyl groups. Enzyme-linked lectin assay was used to screen PS-SGCL against two plant lectins, Glycine max soybean agglutinin and Vicia villosa. The results revealed a carbohydrate density-dependent affinity trend and site-specific glycosylation requirements for high affinity binding to these lectins. Hence, PS-SGCLs provide a platform to systematically elucidate MUC1-lectin binding specificities, which in the long term may provide a rational design for novel inhibitors of MUC1-lectin interactions involved in tumor spread and glycopeptide-based cancer vaccines.
Insights
Synthetic tools were developed to study how MUC1 glycans on cancer cells contribute to metastasis. This research created a novel library to understand MUC1-lectin interactions, aiding cancer vaccine development.
Area of Science:
- Glycoscience
- Cancer Biology
- Synthetic Chemistry
Background:
- Tumor-associated glycans on MUC1 are key in cancer metastasis.
- Epitope heterogeneity on cancer cells hinders understanding of glycan function.
- Lectin interactions with MUC1 glycans are implicated in tumor spread.
Purpose of the Study:
- To design synthetic tools for exploring MUC1 glycan-lectin interactions in metastasis.
- To synthesize a MUC1-derived positional scanning synthetic glycopeptide combinatorial library (PS-SGCL).
- To investigate the role of Tn antigen number and location in MUC1-lectin binding.
Main Methods:
- Synthesis of PS-SGCL using the "tea bag" approach.
- Determination of isokinetic ratios for equimolar incorporation of glycoamino acids.
- Development of an on-resin deprotection protocol for O-acetyl groups.
- Screening of PS-SGCL against soybean agglutinin and Vicia villosa lectins using enzyme-linked lectin assay.
Main Results:
- The study successfully synthesized a novel PS-SGCL with varying Tn antigen.
- Carbohydrate density-dependent affinity trends were observed.
- Site-specific glycosylation was found to be crucial for high-affinity lectin binding.
Conclusions:
- PS-SGCLs offer a platform for systematic elucidation of MUC1-lectin binding specificities.
- Findings may inform the rational design of MUC1-lectin interaction inhibitors.
- This work could lead to the development of novel glycopeptide-based cancer vaccines.
