Label-Free Nanopore Single-Molecule Measurement of Trypsin Activity

Zhou, Shuo; Wang, Liang; Chen, Xiaohan; Guan, Xiyun

doi:10.1021/acssensors.6b00043

Cited by 61 publications

(53 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…0.15 M NaCl ( cis )/3 M NaCl ( trans )), or a greater applied voltage (e.g. +200 mV), as well as other engineered αHL nanopores, as documented in our previous studies .…”

Section: Resultssupporting

confidence: 71%

Nanopore label‐free detection of single‐nucleotide deletion in Baxα/BaxΔ2

et al. 2018

Self Cite

View full text Add to dashboard Cite

Baxα, a key tumor suppressor gene, will not be expressed correctly as a result of single nucleotide mutation in its microsatellite region; Instead, BaxΔ2, an isoform of Baxα, is often produced. In addition, lack of the exon 2 due to an alternative splicing, BaxΔ2 has the same sequence as Baxα except single base deletion from eight continuous guanines (G8) to G7. Most of the currently available methods for Bax∆2 detection are inefficient and time-consuming, and/or require the use of labels or dyes. In this work, we reported a label-free nanopore sensing strategy to differentiate between Baxα and BaxΔ2 with a DNA polymer as a molecular probe based on alternative spliced sequences. Two DNA molecules were designed to selectively detect Baxα and BaxΔ2, respectively. The method was rapid, accurate, and highly sensitive: picomolar concentrations of target nucleic acids could be detected in minutes. Our developed simple and fast nanopore-based detection strategy is not only useful for distinguishing between Baxα and Bax∆2, but also provides a useful tool for detection of other single-base mutations in genetic diagnosis.

show abstract

“…0.15 M NaCl ( cis )/3 M NaCl ( trans )), or a greater applied voltage (e.g. +200 mV), as well as other engineered αHL nanopores, as documented in our previous studies .…”

Section: Resultssupporting

confidence: 71%

Nanopore label‐free detection of single‐nucleotide deletion in Baxα/BaxΔ2

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…They demonstrated LF detection down to 10 n m . Trypsin‐based cleavage studies have also been reported by several groups . Figure C shows nanopore detection of trypsin activity on amyloid beta peptides as detected by a mutated form of the αHL pore (M113F) 7 .…”

Section: Peptide Sensingmentioning

confidence: 67%

The Utility of Nanopore Technology for Protein and Peptide Sensing

Robertson

Reiner

2018

Proteomics

View full text Add to dashboard Cite

Resistive pulse nanopore sensing enables label-free single-molecule analysis of a wide range of analytes. An increasing number of studies have demonstrated the feasibility and usefulness of nanopore sensing for protein and peptide characterization. Nanopores offer the potential to study a variety of protein-related phenomena that includes unfolding kinetics, differences in unfolding pathways, protein structure stability, and free-energy profiles of DNA-protein and RNA-protein binding. In addition to providing a tool for fundamental protein characterization, nanopores have also been used as highly selective protein detectors in various solution mixtures and conditions. This review highlights these and other developments in the area of nanopore-based protein and peptide detection.

show abstract

“…After single connector was inserted and then the translocation signals of R11 or R12 peptide were observed under −50 mV, the trypsin was added and mixed into the cis-chamber while recording the data until the majority of R11 or R12 was digested. [51]…”

Section: Methodsmentioning

confidence: 99%

Nano-channel of viral DNA packaging motor as single pore to differentiate peptides with single amino acid difference

Kang

Wang

et al. 2018

Biomaterials

View full text Add to dashboard Cite

Detection, differentiation, mapping, and sequencing of proteins are important in proteomics for the assessment of cell development such as protein methylation or phosphorylation as well as the diagnosis of diseases including metabolic disorder, mental illness, immunological ailments, and malignant cancers. Nanopore technology has demonstrated the potential for the sequencing or sensing of DNA, RNA, chemicals, or other macromolecules. Due to the diversity of protein in shape, structure and charge and the composition versatility of 20 amino acids, the sequencing of proteins remains challenging. Herein, we report the application of the channel of bacteriophage T7 DNA packaging motor for the differentiation of an assortment of peptides of a single amino acid difference. Explicit fingerprints or signatures were obtained based on current blockage and dwell time of individual peptide. Data from the clear mapping of small proteins after protease digestion suggests the potential of using T7 motor channel for proteomics including protein sequencing.

show abstract

Label-Free Nanopore Single-Molecule Measurement of Trypsin Activity

Cited by 61 publications

References 46 publications

Nanopore label‐free detection of single‐nucleotide deletion in Baxα/BaxΔ2

Nanopore label‐free detection of single‐nucleotide deletion in Baxα/BaxΔ2

The Utility of Nanopore Technology for Protein and Peptide Sensing

Nano-channel of viral DNA packaging motor as single pore to differentiate peptides with single amino acid difference

Contact Info

Product

Resources

About