ArticleNucleic acids research2010
Profiling the selectivity of DNA ligases in an array format with mass spectrometry.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Point-of-care testing ofHeliyon · 2024Article
- Advances in Genotyping Detection of Fragmented Nucleic Acids.Biosensors · 2024Review
- Label-Free Screening of SARS-CoV-2 NSP14 Exonuclease Activity Using SAMDI Mass Spectrometry.SLAS discovery : advancing life sciences R & D · 2021Article
- Structured sequences emerge from random pool when replicated by templated ligation.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Exploring the Ligand Preferences of the PHD1 Domain of Histone Demethylase KDM5A Reveals Tolerance for Modifications of the Q5 Residue of Histone 3.ACS chemical biology · 2021Article
- Article
- Design of glycosylation sites by rapid synthesis and analysis of glycosyltransferases.Nature chemical biology · 2018Article
- Characterization of background noise in capture-based targeted sequencing data.Genome biology · 2017Article
- Structure-activity relationships of the ATP cofactor in ligase-catalysed oligonucleotide polymerisations.Organic & biomolecular chemistry · 2017Article
- Kinetic mechanism and fidelity of nick sealing by Escherichia coli NAD+-dependent DNA ligase (LigA).Nucleic acids research · 2016Article
- A high-throughput assay for the comprehensive profiling of DNA ligase fidelity.Nucleic acids research · 2016Article
- Enzyme-guided DNA Sewing Architecture.Scientific reports · 2015Article
- One-Step Ligation on RNA Amplification for the Detection of Point Mutations.The Journal of molecular diagnostics : JMD · 2015Article
- Article
- Ultrasensitive single-nucleotide polymorphism detection using target-recycled ligation, strand displacement and enzymatic amplification.Nanoscale · 2013Article
- High-throughput screening of small molecule libraries using SAMDI mass spectrometry.ACS combinatorial science · 2011Article
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2 authors.
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Abstract
This article describes a method for the global profiling of the substrate specificities of DNA ligases and illustrates examples using the Taq and T4 DNA ligases. The method combines oligonucleotide arrays, which offer the benefits of high throughput and multiplexed assays, with mass spectrometry to permit label-free assays of ligase activity. Arrays were prepared by immobilizing ternary biotin-tagged DNA substrates to a self-assembled monolayer presenting a layer of streptavidin protein. The array represented complexes having all possible matched and mismatched base pairs at the 3' side of the nick site and also included a number of deletions and insertions at this site. The arrays were treated with ligases and adenosine triphosphate or analogs of the nucleotide triphosphate and then analyzed by matrix-assisted laser desorption-ionization mass spectrometry to determine the yields for both adenylation of the 5'-probe strand and joining of the two probe strands. The resulting activity profiles reveal the basis for specificity of the ligases and also point to strategies that use ATP analogs to improve specificity. This work introduces a method that can be applied to profile a broad range of enzymes that operate on nucleic acid substrates.
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