Surface-Mediated Protein Unfolding as a Search Process for Denaturing Sites.
Surface-Mediated Protein Unfolding as a Search Process for Denaturing Sites.
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DOI：
10.1021/acsnano.5b05787
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发表时间：
2016-01
期刊：
影响因子：
17.1
通讯作者：
James S. Weltz;D. K. Schwartz;Joel L. Kaar
中科院分区：
文献类型：
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作者：
James S. Weltz;D. K. Schwartz;Joel L. Kaar
表面诱导的蛋白质变性对于开发对生物分子具有抗性和/或无害的材料具有重要意义。在这里，我们研究了溶菌酶（T4 L）在熔融石英（FS）上的解折叠机制，使用单分子方法，提供了直接洞察变性的原因。T4 L的展开通过Förster共振能量转移进行监测，同时跟踪单个分子在固溶体界面处的吸附、扩散和解吸。高通量单分子分析的结果表明，FS上的T4 L的展开是由表面扩散介导的，并发生在孤立的纳米级网站，这是相对罕见的，不同于大多数的表面。这些观察结果表明，表面介导的蛋白质解折叠是一个搜索过程，是基于对变性位点的蛋白质的探索。最终，这些发现对蛋白质相容表面的设计具有重要意义。
Surface-induced protein denaturation has important implications for the development of materials that are resistant and/or innocuous to biomolecules. Here, we studied the mechanism of lysozyme (T4L) unfolding on fused silica (FS) using single-molecule methods that provided direct insight into the cause of denaturation. Unfolding of T4L was monitored by Förster resonance energy transfer while simultaneously tracking the adsorption, diffusion, and desorption of individual molecules at the solid-solution interface. Results of high-throughput single-molecule analysis suggested that the unfolding of T4L on FS was mediated by surface diffusion and occurred on isolated nanoscale sites, which were relatively rare and distinct from the majority of the surface. These observations suggest that surface-mediated protein unfolding is a search process that is based on the exploration for denaturing sites by the protein. Ultimately, these findings have important implications for the design of protein-compatible surfaces.