Single-molecule resolution of macromolecules with nanopore devices

Advances in Colloid and Interface Science, Journal Year: 2025, Volume and Issue: unknown, P. 103417 - 103417

Published: Jan. 1, 2025

Language: Английский

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Aerolysin Nanopore Electrochemistry

Accounts of Chemical Research, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 28, 2025

ConspectusIons are the crucial signaling components for living organisms. In cells, their transportation across pore-forming membrane proteins is vital regulating physiological functions, such as generating ionic current signals in response to target molecule recognition. This ion transport affected by confined interactions and local environments within protein pore. Therefore, can efficiently transduce characteristics of each into ion-transport-mediated with high sensitivity. Inspired nature, various pores have been developed high-throughput label-free nanopore sensors single-molecule detection, enabling rapid accurate readouts. particular, aerolysin, a key virulence factor Aeromonas hydrophila, exhibits sensitivity fingerprints detecting subtle differences sequence, conformation, structure DNA, proteins, polypeptides, oligosaccharides, other molecules. Aerolysin features cap that approximately 14 nm wide on cis side central pore about 10 long minimum diameter around 1 nm. Its lumen, 11 charged rings at two entrances neutral amino acids between, facilitates dwelling single analyte characteristic enables rich between well-defined residues analyte. As result, signal offers unique molecular fingerprint, extending beyond traditional volume exclusion model sensing. 2006, aerolysin was first reported discriminate conformational peptides, opening door rapidly growing field electrochemistry. Over years, mutant nanopores emerged, associated advanced instrumentation data analysis algorithms, simultaneous identification over 30 targets number still increasing. electrochemistry particular allows time-resolved qualitative quantitative ranging from DNA sequencing, proteomics, enzyme kinetics, reactions potential clinical diagnostics. Especially, feasibility dynamic would revolutionize omics studies level, paving way promising temporal omics. Despite success this approach so far, it remains challenging understand how correlate distinguishable signatures. Recent attempts added correction terms account variations mobility caused Account, we revisit origin blockade induced molecules inside nanopore. We highlight contributions noncovalent sensing ability through corrected conductance model. Account then describes design interaction networks nanopore, including electrostatic, hydrophobic, hydrogen-bonding, cation−π, ion–charged acid interactions, ultrasensitive biomolecular quantification. Finally, provide an outlook further understanding network improving manipulating fine-tuning toward broad range practical applications.

Language: Английский

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Emerging protein sequencing technologies: proteomics without Mass spectrometry?

Expert Review of Proteomics, Journal Year: 2025, Volume and Issue: unknown

Published: March 19, 2025

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been a leading method for proteomics thirty years. Advantages provided by LC-MS/MS are offset significant disadvantages, including cost. Recently, several non-mass spectrometric methods have emerged, but little information is available about their capacity to analyze the complex mixtures routine spectrometry. We review recent non-mass-spectrometric sequencing proteins and peptides, using nanopores, degradation, reverse translation, short-epitope mapping, with comments on bioinformatics challenges, fundamental limitations, areas where new technologies will be more or less competitive LC-MS/MS. In addition conventional literature searches, instrument vendor websites, patents, webinars, preprints were also consulted give up-to-date picture. Many promising. However, demonstrations that they outperform in terms of peptides identified not yet published, astute observers note important especially relating dynamic range single-molecule measurements mixtures. Still, even if performance emerging proves inferior LC-MS/MS, low cost could create different kind revolution: dramatic increase number biology laboratories engaging forms research.

Language: Английский

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Post‐Treatment Apical Periodontitis: Endodontic Infection Contributes to Circulating Microbial DNA

Australian Endodontic Journal, Journal Year: 2025, Volume and Issue: unknown

Published: March 27, 2025

ABSTRACT This study investigated the impact of post‐treatment apical periodontitis on microbial DNA in patients' blood circulation. Samples from root apex, periapical lesion and peripheral 11 patients, along with nine healthy controls, were analysed using next‐generation sequencing. The composition, diversity linear discriminant analysis effect size analysed. SourceTracker was applied to trace microorganisms blood. Proteobacteria dominated lesions, while Actinobacteria prevailed apex. Patients exhibited higher but reduced α‐diversity apex compared other sites. Moreover, Bacteroides , as well Aliihoeflea Oceanicaulis Comamonas Shigella Acinetobacter showed increased relative abundance patients. Additionally, contributed 26% 76% for bacteria. These findings suggested unique translocation

Language: Английский

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