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Autori principali: Murphy, Ian, Bobilev, Keren, Hayakawa, Daichi, Ikonen, Eden, Videbæk, Thomas E., Dalal, Shibani, Ahmed, Wylie W., Ross, Jennifer L., Rogers, W. Benjamin
Natura: Preprint
Pubblicazione: 2024
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Accesso online:https://arxiv.org/abs/2409.03040
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author Murphy, Ian
Bobilev, Keren
Hayakawa, Daichi
Ikonen, Eden
Videbæk, Thomas E.
Dalal, Shibani
Ahmed, Wylie W.
Ross, Jennifer L.
Rogers, W. Benjamin
author_facet Murphy, Ian
Bobilev, Keren
Hayakawa, Daichi
Ikonen, Eden
Videbæk, Thomas E.
Dalal, Shibani
Ahmed, Wylie W.
Ross, Jennifer L.
Rogers, W. Benjamin
contents Attaching enzymes to nanostructures has proven useful to the study of enzyme functionality under controlled conditions and has led to new technologies. Often, the utility and interest of enzyme-tethered nanostructures lie in how the enzymatic activity is affected by how the enzymes are arranged in space. Therefore, being able to conjugate enzymes to nanostructures while preserving the enzymatic activity is essential. In this paper, we present a method to conjugate single-stranded DNA to the enzyme urease while maintaining enzymatic activity. We show evidence of successful conjugation and quantify the variables that affect the conjugation yield. We also show that the enzymatic activity is unchanged after conjugation compared to the enzyme in its native state. Finally, we demonstrate the tethering of urease to nanostructures made using DNA origami with high site-specificity. Decorating nanostructures with enzymatically-active urease may prove to be useful in studying, or even utilizing, the functionality of urease in disciplines ranging from biotechnology to soft-matter physics. The techniques we present in this paper will enable researchers across these fields to modify enzymes without disrupting their functionality, thus allowing for more insightful studies into their behavior and utility.
format Preprint
id arxiv_https___arxiv_org_abs_2409_03040
institution arXiv
publishDate 2024
record_format arxiv
spellingShingle A method for site-specifically tethering the enzyme urease to DNA origami with sustained activity
Murphy, Ian
Bobilev, Keren
Hayakawa, Daichi
Ikonen, Eden
Videbæk, Thomas E.
Dalal, Shibani
Ahmed, Wylie W.
Ross, Jennifer L.
Rogers, W. Benjamin
Biological Physics
Attaching enzymes to nanostructures has proven useful to the study of enzyme functionality under controlled conditions and has led to new technologies. Often, the utility and interest of enzyme-tethered nanostructures lie in how the enzymatic activity is affected by how the enzymes are arranged in space. Therefore, being able to conjugate enzymes to nanostructures while preserving the enzymatic activity is essential. In this paper, we present a method to conjugate single-stranded DNA to the enzyme urease while maintaining enzymatic activity. We show evidence of successful conjugation and quantify the variables that affect the conjugation yield. We also show that the enzymatic activity is unchanged after conjugation compared to the enzyme in its native state. Finally, we demonstrate the tethering of urease to nanostructures made using DNA origami with high site-specificity. Decorating nanostructures with enzymatically-active urease may prove to be useful in studying, or even utilizing, the functionality of urease in disciplines ranging from biotechnology to soft-matter physics. The techniques we present in this paper will enable researchers across these fields to modify enzymes without disrupting their functionality, thus allowing for more insightful studies into their behavior and utility.
title A method for site-specifically tethering the enzyme urease to DNA origami with sustained activity
topic Biological Physics
url https://arxiv.org/abs/2409.03040