Tuning Protein Discrimination Through Altering the Sampling Interface Formed between the Analyte and the OmpG Nanopore,ACS Sensors

当前位置： X-MOL 学术 › ACS Sens. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Tuning Protein Discrimination Through Altering the Sampling Interface Formed between the Analyte and the OmpG Nanopore
ACS Sensors ( IF 8.2 ) Pub Date : 2021-02-18 , DOI: 10.1021/acssensors.0c02580
Monifa A Fahie _{1,

2} , Jonathan Candido ₂ , Gisele Andree ₂ , Min Chen _{1,

2}

Affiliation

Nanopore sensors capable of distinguishing homologous protein analytes are highly desirable tools for proteomics research and disease diagnostics. Recently, an engineered outer membrane protein G (OmpG) nanopore with a high-affinity ligand attached to a gating loop 6 showed specificity for distinguishing homologous proteins in complex mixtures. Here, we report the development of OmpG nanopores with the other six loops used as the anchoring point to host an affinity ligand for protein sensing. We investigated how the analyte binding to the affinity ligand located at different loops affects the detection sensitivity, selectivity, and specificity. Our results reveal that analytes weakly attracted to the OmpG nanopore surface are only detectable when the ligand is tethered to loop 6. In contrast, protein analytes that form a strong interaction with the OmpG surface via electrostatic attractions are distinguishable by all seven OmpG nanopore constructs. In addition, the same analyte can generate distinct binding signals with different OmpG nanopore constructs. The ability to exploit all seven OmpG loops will aid the design of a new generation of OmpG sensors with increased sensitivity, selectivity, and specificity for biomarker sensing.

中文翻译：

通过改变分析物和 OmpG 纳米孔之间形成的采样界面来调整蛋白质识别

能够区分同源蛋白质分析物的纳米孔传感器是蛋白质组学研究和疾病诊断的理想工具。最近，工程化外膜蛋白 G (OmpG) 纳米孔与连接到门控环 6 的高亲和力配体显示出区分复杂混合物中同源蛋白质的特异性。在这里，我们报告了 OmpG 纳米孔的发展，其他六个环用作锚定点以承载用于蛋白质传感的亲和配体。我们研究了分析物与位于不同环的亲和配体的结合如何影响检测灵敏度、选择性和特异性。我们的结果表明，只有当配体连接到环 6 时，才可检测到被 OmpG 纳米孔表面弱吸引的分析物。相比之下，通过静电吸引与 OmpG 表面形成强相互作用的蛋白质分析物可通过所有七种 OmpG 纳米孔结构来区分。此外，相同的分析物可以与不同的 OmpG 纳米孔结构产生不同的结合信号。利用所有七个 OmpG 回路的能力将有助于设计具有更高灵敏度、选择性和特异性的生物标志物传感的新一代 OmpG 传感器。

更新日期：2021-03-26

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11