Wood Amanda M, Wardzala Casia L, Kramer Jessica R
Department of Biomedical Engineering, University of Utah Salt Lake City UT 84112 USA
RSC Chem Biol. 2025 Jul 14. doi: 10.1039/d5cb00111k.
Mucin proteins are essential for life but are challenging to study due to their complex glycosylation patterns. Synthetic mimics have become vital tools for understanding and modulating the roles of mucins in human health and disease. These materials also have diverse biomedical applications as lubricants and anti-infectives, in vaccine formulations, and more. We developed a chemoenzymatic approach to prepare polypeptide-based synthetic mucins displaying a variety of glycans with native linkages and orientations. By combining the polymerization of glycosylated amino acid -carboxyanhydrides with enzymatic sialylation and fucosylation, we produced a tunable panel of synthetic mucins. These polymers were recognized by natural glycan-binding and glycan-degrading enzymes, providing insights into the structural preferences of these proteins. Glycan- and linkage-dependent effects on proteolysis were observed. Further, investigation of the influence of glycans on peptide backbone secondary structure revealed that both sialylation and linkage at Ser Thr have profound effects on hierarchical conformation. Overall, our methodology offers versatile tools for exploring the diverse glycobiology of mucins.
粘蛋白对生命至关重要，但由于其复杂的糖基化模式，研究起来颇具挑战性。合成模拟物已成为理解和调节粘蛋白在人类健康与疾病中作用的重要工具。这些材料在作为润滑剂和抗感染剂、疫苗制剂等方面也有多种生物医学应用。我们开发了一种化学酶法来制备基于多肽的合成粘蛋白，这些粘蛋白展示出具有天然连接和取向的多种聚糖。通过将糖基化氨基酸 - 羧基酐的聚合与酶促唾液酸化和岩藻糖基化相结合，我们制备了一组可调节的合成粘蛋白。这些聚合物能被天然的聚糖结合酶和聚糖降解酶识别，从而为了解这些蛋白质的结构偏好提供了线索。观察到了聚糖和连接对蛋白水解的影响。此外，对聚糖对肽主链二级结构影响的研究表明，丝氨酸和苏氨酸处的唾液酸化和连接对分级构象都有深远影响。总体而言,我们的方法为探索粘蛋白多样的糖生物学提供了通用工具。
