Resolving Sulfation Posttranslational Modifications on a Peptide Hormone using Nanopores

ACS Nano, Год журнала: 2024, Номер 18(42), С. 28999 - 29007

Опубликована: Окт. 10, 2024

Peptide hormones are decorated with post-translational modifications (PTMs) that crucial for receptor recognition. Tyrosine sulfation on plant peptide is, example, essential growth and development. Measuring the occurrence position of sulfotyrosine however, compromised by major technical challenges during isolation detection. Nanopores can sensitively detect protein PTMs at single-molecule level. By translocating PTM variants pentapeptide hormone phytosulfokine (PSK) through a nanopore, we here demonstrate accurate identification phosphorylation two tyrosine residues PSK. Sulfation be clearly detected distinguished (>90%) from same residue. Moreover, presence or absence close-by accurately determined (>96% accuracy). Our findings extraordinary sensitivity nanopore measurements, providing powerful tool identifying position-specific promising wider applications to identify PTMs.

Язык: Английский

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Obtaining Narrow Distributions of Single-Molecule Peptide Signals Enables Sensitive Peptide Discrimination with α-Hemolysin Nanopores

Journal of the American Chemical Society, Год журнала: 2025, Номер unknown

Опубликована: Март 10, 2025

Biological nanopore technology has emerged as a promising tool for analyzing peptides and post-translational modifications at the single-molecule level. However, broader application is currently limited by partial separation of low-throughput, mainly due to nonuniform peptide signals detected nanopores. Narrowing signal distribution crucial improving nanopore's sensing ability but remains bottleneck. Here, we demonstrate that capturing with electrophoretic force against electroosmotic flow can provoke more uniform blockades in α-hemolysin By using buffers 2 M KCl pH 3.8, obtain most signals, which may be correlated shape, linearization, actual dwelling position peptides. Five acetylation phosphorylation, including isomeric peptides, readily separated from each other. The citrullination replacement arginine β-hydroxybutyrylation modification another sequence are also discriminated mixture. A series different compositions induced when they were analyzed our method. Our work presents an efficient approach optimize analysis

Язык: Английский

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Eliminating the Interference of Neighboring Nucleobases in Aerolysin for Nanopore Sequencing

ACS Sensors, Год журнала: 2025, Номер unknown

Опубликована: Апрель 8, 2025

Biological nanopores have revolutionized DNA sequencing with their incredible advantages of long reads, high throughput, portability, and low material requirement. Despite numerous improvements, base calling remains challenging due to the influence neighboring nucleobases at reading site. One direction development is exploring or designing new improve accuracy. Aerolysin emerging as a powerful candidate, which can identify subtle differences detected molecules. However, not many studies focus on translating these into sequencing. Here, we used streptavidin immobilize ssDNA inside an engineered aerolysin channel, allows us investigate its sensing regions, bases influence. By increasing voltage, eliminate reduce k-mer size one nucleobase. These findings hold great potential for improving accuracy nanopore well nanopore-based other polymers.

Язык: Английский

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Controlled Translocation of Proteins through a Biological Nanopore for Single-Protein Fingerprint Identification

Nano Letters, Год журнала: 2024, Номер 24(44), С. 14118 - 14124

Опубликована: Окт. 24, 2024

After the successful sequencing of nucleic acids, nanopore technology has now been applied to proteins. Recently, it demonstrated that an electro-osmotic flow can be used induce transport unraveled polypeptides across nanopores. Polypeptide translocation, however, is too fast for accurate reading its amino acid compositions. Here, we show introduction hydrophobic residues into lumen reduces protein translocation speed. Importantly, a tyrosine at entry and isoleucine β-barrel reduced speed ∼10 acids/millisecond while keeping relatively large ionic current, crucial component identification. These nanopores showed unique features within their current signatures, which may pave way toward fingerprinting using

Язык: Английский

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Single-molecule protein sequencing with nanopores

Nature Reviews Bioengineering, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 21, 2024

Язык: Английский

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