Aggregation and Oligomerization Characterization of ß-Lactoglobulin Protein Using a Solid-State Nanopore Sensor.
Aggregation and Oligomerization Characterization of ß-Lactoglobulin Protein Using a Solid-State Nanopore Sensor.
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DOI：
10.3390/s24010081
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发表时间：
2023-12-22
期刊：
影响因子：
--
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Acharjee MC;Ledden B;Thomas B;He X;Messina T;Giurleo J;Talaga D;Li J
蛋白质聚集与许多慢性和破坏性的神经退行性人类疾病有关，并且与衰老密切相关。这项工作表明，蛋白质聚集和寡聚化可以通过固态纳米孔方法在单分子水平上进行评估。氮化硅纳米孔传感器用于表征模型蛋白β-乳球蛋白变体A（βLGa）的淀粉样蛋白生成和天然状态寡聚化。纳米孔测量的结果针对原子力显微镜（AFM）和动态光散射（DLS）数据进行了验证，比较了不同阶段相同样品的βLGa聚集。通过线性和环状双链DNA的校准，本研究估计了淀粉样纤维的长度和直径，βLGa聚集体的数量及其分布。纳米孔结果与DLS和AFM数据一致，并在单个蛋白质分子组装体的水平上提供了额外的见解。作为纳米孔技术的进一步证明，在高（2 M KCl）和低（0.1 M KCl）离子强度下测量了pH 4.6下βLGa自缔合和聚集随温度的变化。这项研究强调了使用固态纳米孔方法分析蛋白质聚集的优点和局限性。
Protein aggregation is linked to many chronic and devastating neurodegenerative human diseases and is strongly associated with aging. This work demonstrates that protein aggregation and oligomerization can be evaluated by a solid-state nanopore method at the single molecule level. A silicon nitride nanopore sensor was used to characterize both the amyloidogenic and native-state oligomerization of a model protein ß-lactoglobulin variant A (βLGa). The findings from the nanopore measurements are validated against atomic force microscopy (AFM) and dynamic light scattering (DLS) data, comparing βLGa aggregation from the same samples at various stages. By calibrating with linear and circular dsDNA, this study estimates the amyloid fibrils’ length and diameter, the quantity of the βLGa aggregates, and their distribution. The nanopore results align with the DLS and AFM data and offer additional insight at the level of individual protein molecular assemblies. As a further demonstration of the nanopore technique, βLGa self-association and aggregation at pH 4.6 as a function of temperature were measured at high (2 M KCl) and low (0.1 M KCl) ionic strength. This research highlights the advantages and limitations of using solid-state nanopore methods for analyzing protein aggregation.
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影响因子：
2.9
作者：
Uplinger, James;Thomas, Brian;Rollings, Ryan;Fologea, Daniel;McNabb, David;Li, Jiali
通讯作者：
Li, Jiali
影响因子：
14.9
作者：
Dorvel B;Sigalov G;Zhao Q;Comer J;Dimitrov V;Mirsaidov U;Aksimentiev A;Timp G
通讯作者：
Timp G
影响因子：
3
作者：
Pulawski, Wojciech;Ghoshdastider, Umesh;Andrisano, Vincenza;Filipek, Slawomir
通讯作者：
Filipek, Slawomir
影响因子：
3.5
作者：
Rollings R;Graef E;Walsh N;Nandivada S;Benamara M;Li J
通讯作者：
Li J
影响因子：
1.6
作者：
Li J;Fologea D;Rollings R;Ledden B
通讯作者：
Ledden B