Chemical, Thermal, and Electric Field Induced Unfolding of Single Protein Molecules Studied Using Nanopores
Chemical, Thermal, and Electric Field Induced Unfolding of Single Protein Molecules Studied Using Nanopores
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DOI：
10.1021/ac2001725
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发表时间：
2011-07-01
影响因子：
7.4
通讯作者：
Kim, Min Jun
中科院分区：
文献类型：
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作者：
Freedman, Kevin J.;Jurgens, Maike;Kim, Min Jun
最近，单分子实验技术在研究实验室和工业环境中的许多应用中都显示出了巨大的兴趣。尽管存在许多单分子技术，但纳米孔平台可能是较受欢迎的技术之一，因为它能够充当生物大分子的分子传感器。例如，纳米孔为探测蛋白质的各种性质提供了一种独特的新方法，并可结合现有技术有助于阐明关键的生物物理信息。在本研究中，检测到了各种形式的牛血清白蛋白(BSA)，包括热复性BSA、尿素变性的BSA，以及在提高电场强度(有或没有尿素)下检测到的多种形式的BSA。我们还为这些状态中的每一个提供了排除的体积测量，这些状态通常由于未知和不稳定的蛋白质构象而难以获得。
Single-molecule experimental techniques have recently shown to be of significant interest for use in numerous applications in both the research laboratory and industrial settings. Although many single-molecule techniques exist, the nanopore platform is perhaps one of the more popular techniques due to its ability to act as a molecular sensor of biological macromolecules. For example, nanopores offer a unique, new method for probing various properties of proteins and can contribute to elucidating key biophysical information in conjunction with existing techniques. In the present study, various forms of bovine serum albumin (BSA) are detected including thermally refolded BSA, urea-denatured BSA, and multiple forms of BSA detected at elevated electric field strengths (with and without urea). We also provide excluded volume measurements for each of these states that normally are difficult to obtain due to unknown and unstable protein conformations.