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Related Concept Videos

Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

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Related Experiment Video

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Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor
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Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor

Published on: March 21, 2018

A selective adenosine sensor derived from a triplex DNA aptamer.

Mayurbhai Patel1, Avishek Dutta, Haidong Huang

  • 1Department of Chemistry and Environmental Science, New Jersey Institute of Technology, Newark, 07102, USA.

Analytical and Bioanalytical Chemistry
|May 7, 2011
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel adenosine aptamer sensor using triplex DNA for enhanced affinity and selectivity. This sensor offers a sensitive and specific method for detecting adenosine concentrations.

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Area of Science:

  • Biotechnology
  • Nucleic Acid Chemistry
  • Biosensors

Background:

  • Traditional aptamer development relies on SELEX, which can be inefficient for small molecules.
  • Duplex DNA with abasic sites offers a scaffold for rational aptamer design.
  • Triplex DNA structures present an alternative scaffold for aptamer development.

Purpose of the Study:

  • To rationally design a selective adenosine aptamer sensor using a triplex DNA scaffold.
  • To investigate the potential of abasic site-containing triplex DNA as an aptamer scaffold.
  • To develop a fluorescence-based sensor for adenosine detection.

Main Methods:

  • Rational design of a triplex DNA aptamer incorporating a furano-dU modification.
  • Characterization of the aptamer-adenosine binding affinity using dissociation constant measurements.
  • Assessment of sensor selectivity against related nucleosides and nucleotides.
  • Determination of the sensor's detection limit and quantification range.

Main Results:

  • The triplex DNA aptamer exhibited a high affinity for adenosine with a dissociation constant of 400 nM.
  • Adenosine binding resulted in a 40% quenching of furano-dU fluorescence.
  • The sensor demonstrated high selectivity for adenosine over uridine, cytidine, guanosine, ATP, and AMP.
  • The sensor achieved a detection limit of approximately 50 nM and could quantify adenosine from 50 nM to 2 μM.

Conclusions:

  • Abasic site-containing triplex DNA serves as an effective scaffold for rational aptamer design.
  • The developed adenosine aptamer sensor is selective, sensitive, and exhibits improved affinity compared to SELEX-derived aptamers.
  • This novel sensor provides a valuable tool for quantifying adenosine in biological samples.