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Related Experiment Video

Updated: Jun 4, 2026

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1
08:17

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1

Published on: April 16, 2021

A high-throughput assay for DNA deaminases.

Meng Wang1, Cristina Rada, Michael S Neuberger

  • 1Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.

Methods in Molecular Biology (Clifton, N.J.)
|March 4, 2011
PubMed
Summary

Researchers adapted bacterial papillation assays to measure the DNA mutator activity of AID/APOBEC proteins. This high-throughput screening method can identify enhanced AID variants and novel DNA deaminases.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • The AID/APOBEC family comprises DNA deaminases that catalyze cytosine to uracil conversion.
  • This deamination activity functions as a DNA mutator mechanism.

Purpose of the Study:

  • To adapt bacterial papillation assays for monitoring AID/APOBEC mutator activity.
  • To establish a high-throughput screen for identifying AID variants with enhanced specific activity.
  • To explore the use of papillation assays for discovering novel DNA deaminases.

Main Methods:

  • Adaptation of bacterial papillation assays.
  • High-throughput screening of AID/APOBEC proteins.
  • Monitoring DNA deamination activity.

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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis

Published on: April 22, 2022

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Last Updated: Jun 4, 2026

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1
08:17

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1

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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis

Published on: April 22, 2022

Main Results:

  • Demonstrated the utility of bacterial papillation assays in measuring AID/APOBEC mutator activity.
  • Successfully employed the assay as a high-throughput screen to identify AID variants with increased specific activity.

Conclusions:

  • Bacterial papillation assays are a versatile tool for studying DNA deaminases.
  • The developed screening method facilitates the discovery of novel DNA deaminases and improved AID variants.