An Ultrastable and Dense Single-Molecule Click Platform for Sensing Protein–Deoxyribonucleic Acid Interactions

Emiel W.A. Visser; Jovana Miladinovic; Joshua N. Milstein

doi:10.1002/smtd.202001180

An Ultrastable and Dense Single-Molecule Click Platform for Sensing Protein–Deoxyribonucleic Acid Interactions

Emiel W.A. Visser (Corresponding author), Jovana Miladinovic, Joshua N. Milstein (Corresponding author)

Molecular Biosensing

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

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An ultrastable, highly dense single-molecule assay ideal for observing protein–DNA interactions is demonstrated. Stable click tethered particle motion leverages next generation click-chemistry to achieve an ultrahigh density of surface tethered reporter particles, and has low non-specific interactions, is stable at elevated temperatures to at least 45 °C, and is compatible with Mg²⁺, an important ionic component of many regulatory protein–DNA interactions. Prepared samples remain stable, with little degradation, for >6 months in physiological buffers. These improvements enable the authors to study previously inaccessible sequence and temperature-dependent effects on DNA binding by the bacterial protein, histone-like nucleoid-structuring protein, a global transcriptional regulator found in Escherichia coli. This greatly improved assay can directly be translated to accelerate existing tethered particle-based, single-molecule biosensing applications.

Originele taal-2	Engels
Artikelnummer	2001180
Aantal pagina's	6
Tijdschrift	Small Methods
Volume	5
Nummer van het tijdschrift	5
Vroegere onlinedatum	23 feb. 2021
DOI's	https://doi.org/10.1002/smtd.202001180
Status	Gepubliceerd - 12 mei 2021

Financiering

The authors thank Paul Piunno for his helpful discussions on optimizing the APTES functionalization, William Navarre for providing the H‐NS proteins, and Marie Elliot and Xiafei Zhang for providing the natural DNA templates from . This project has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No. 796345, and from a National Sciences and Engineering Research Council of Canada Discovery Grant (RGPIN‐2019‐06520). Streptomyces venezuelae

Financiers	Financiernummer
European Union’s Horizon Europe research and innovation programme
Natural Sciences and Engineering Research Council of Canada	RGPIN‐2019‐06520
European Union’s Horizon Europe research and innovation programme	796345

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Small Methods - An Ultrastable and Dense Single-Molecule Click Platform for Sensing Protein-Deoxyribonucleic Acid interactionsDrukproef, 504 KBLicentie: CC BY-NC

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abstract = "An ultrastable, highly dense single-molecule assay ideal for observing protein–DNA interactions is demonstrated. Stable click tethered particle motion leverages next generation click-chemistry to achieve an ultrahigh density of surface tethered reporter particles, and has low non-specific interactions, is stable at elevated temperatures to at least 45 °C, and is compatible with Mg2+, an important ionic component of many regulatory protein–DNA interactions. Prepared samples remain stable, with little degradation, for >6 months in physiological buffers. These improvements enable the authors to study previously inaccessible sequence and temperature-dependent effects on DNA binding by the bacterial protein, histone-like nucleoid-structuring protein, a global transcriptional regulator found in Escherichia coli. This greatly improved assay can directly be translated to accelerate existing tethered particle-based, single-molecule biosensing applications.",

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An Ultrastable and Dense Single-Molecule Click Platform for Sensing Protein–Deoxyribonucleic Acid Interactions. / Visser, Emiel W.A. (Corresponding author); Miladinovic, Jovana; Milstein, Joshua N. (Corresponding author).
In: Small Methods, Vol. 5, Nr. 5, 2001180, 12.05.2021.

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

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An Ultrastable and Dense Single-Molecule Click Platform for Sensing Protein–Deoxyribonucleic Acid Interactions

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