Genetic diversity leads to protein robustness, adaptability, and failure. Some sequence variants are structurally robust but functionally disturbed because mutations bring the protein onto unfolding/refolding routes resulting in misfolding diseases (e.g., Parkinson). We assume dynamic perturbations introduced by mutations foster the alternative unfolding routes and test this possibility by comparing the unfolding dynamics of the heat-labile enterotoxin B pentamers and the cholera toxin B pentamers, two pentamers structurally and functionally related and robust to 17 sequence variations. The B-subunit thermal unfolding dynamics are monitored by broadband dielectric spectroscopy in nanoconfined and weakly hydrated conditions. Distinct dielectric signals reveal the different B-subunits unfolding dynamics. Combined with network analyses, the experiments pinpoint the role of three mutations A1T, E7D, and E102A, in diverting LTB₅ to alternative unfolding routes that protect LTB₅ from dissociation. Altogether, the methodology diagnoses dynamics faults that may underlie functional disorder, drug resistance, or higher virulence of sequence variants.

遗传多样性导致蛋白质的稳健性、适应性和失败。一些序列变体结构稳健但功能受到干扰，因为突变使蛋白质进入解折叠/重折叠途径，导致错误折叠疾病（例如，帕金森）。我们假设突变引入的动态扰动促进了替代的展开途径，并通过比较热不稳定肠毒素 B 五聚体和霍乱毒素 B 五聚体的展开动力学来测试这种可能性，这两种五聚体在结构和功能上相关并且对 17 个序列变化具有鲁棒性。在纳米限制和弱水合条件下，通过宽带介电光谱监测 B 亚基热展开动力学。不同的介电信号揭示了不同的 B 亚基展开动力学。结合网络分析，₅到保护 LTB ₅免于解离的替代展开路线。总而言之，该方法诊断可能导致功能障碍、耐药性或序列变体更高毒力的动力学故障。