Synthetic chaperones are stimuli-responsive materials that facilitate the refolding of denatured proteins in their native form and release refolded proteins in solution in the presence of external stimuli. Thermoresponsive hydrogels are a class of synthetic chaperones that require heat as an external stimulus to refold the denatured proteins into their biologically active conformations; however, the mechanism by which these hydrogels participate in protein refolding mechanism remains unclear. In this study, we explore the role of physical and structural changes in the hydrogel matrix that may participate in protein refolding efficacies of thermally responsive poly(vitamin B5 analogous methacrylamide) poly(B5AMA) hydrogels. Poly(B5AMA) hydrogels of different net charges, hydrophobicity, and cross-linking densities are synthesized by radical polymerization method and are evaluated for the restoration of enzymatic activity of thermally denatured enzymes (lysozyme and carbonic anhydrase) as a function of temperature and time and for the presence of residual water in the hydrogel architecture. The hydrogels with promising protein renaturation efficacies are further evaluated for their interactions with denatured proteins, and the role of thermal flexibility of the hydrogel matrix in protein refolding capabilities is elucidated.

中文翻译：

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阐明水凝胶的热柔韧性在蛋白质重折叠中的作用

合成伴侣是刺激响应材料，可促进变性蛋白质以其天然形式重新折叠，并在存在外部刺激的情况下在溶液中释放重新折叠的蛋白质。热响应性水凝胶是一类合成伴侣，需要热量作为外部刺激，将变性蛋白质重新折叠成具有生物活性的构象；然而，这些水凝胶参与蛋白质重折叠机制的机制仍不清楚。在这项研究中，我们探讨了水凝胶基质中物理和结构变化的作用，这些变化可能参与热响应性聚（维生素 B5 类似甲基丙烯酰胺）聚 (B5AMA) 水凝胶的蛋白质重折叠功效。不同净电荷、疏水性的聚（B5AMA）水凝胶，通过自由基聚合方法合成交联密度和交联密度，并评估作为温度和时间函数的热变性酶（溶菌酶和碳酸酐酶）的酶活性的恢复以及水凝胶结构中残留水的存在。进一步评估了具有前景的蛋白质复性功效的水凝胶与变性蛋白质的相互作用，并阐明了水凝胶基质的热柔韧性在蛋白质重折叠能力中的作用。