Literature DB >> 33877599

Indirect Detection of Ligand Binding by Thermal Melt Analysis.

Joseph Shaw1, Christopher Stubbs2.   

Abstract

A thermal shift assay (TSA) involves measuring the effect of a compound on the thermal stability of a protein as an indirect measure of ligand binding. In this chapter, we provide a protocol for a conventional TSA with recombinant/purified proteins using differential scanning fluorimetry (DSF), followed by a protocol for a Cellular Thermal Shift Assay (CETSA®), which measures the soluble cellular protein remaining after a transient heat shock of live cells to detect intracellular ligand binding.

Keywords:  CETSA; Cellular Thermal Shift Assay; DSF; Differential scanning fluorimetry; Ligand binding; TSA; Target engagement; Thermal shift assay

Year:  2021        PMID： 33877599     DOI： 10.1007/978-1-0716-1197-5_8

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN： 1064-3745


  12 in total

1.  Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery.

Authors:  Mei-Chu Lo; Ann Aulabaugh; Guixian Jin; Rebecca Cowling; Jonathan Bard; Michael Malamas; George Ellestad
Journal:  Anal Biochem       Date:  2004-09-01       Impact factor: 3.365

2.  The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability.

Authors:  Frank H Niesen; Helena Berglund; Masoud Vedadi
Journal:  Nat Protoc       Date:  2007       Impact factor: 13.491

3.  Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay.

Authors:  Daniel Martinez Molina; Rozbeh Jafari; Marina Ignatushchenko; Takahiro Seki; E Andreas Larsson; Chen Dan; Lekshmy Sreekumar; Yihai Cao; Pär Nordlund
Journal:  Science       Date:  2013-07-05       Impact factor: 47.728

4.  A three-stage biophysical screening cascade for fragment-based drug discovery.

Authors:  Ellene H Mashalidis; Paweł Śledź; Steffen Lang; Chris Abell
Journal:  Nat Protoc       Date:  2013-10-24       Impact factor: 13.491

5.  The cellular thermal shift assay for evaluating drug target interactions in cells.

Authors:  Rozbeh Jafari; Helena Almqvist; Hanna Axelsson; Marina Ignatushchenko; Thomas Lundbäck; Pär Nordlund; Daniel Martinez Molina
Journal:  Nat Protoc       Date:  2014-08-07       Impact factor: 13.491

6.  The use of AlphaScreen technology in HTS: current status.

Authors:  Richard M Eglen; Terry Reisine; Philippe Roby; Nathalie Rouleau; Chantal Illy; Roger Bossé; Martina Bielefeld
Journal:  Curr Chem Genomics       Date:  2008-02-25

7.  Microscale fluorescent thermal stability assay for membrane proteins.

Authors:  Alexander I Alexandrov; Mauro Mileni; Ellen Y T Chien; Michael A Hanson; Raymond C Stevens
Journal:  Structure       Date:  2008-03       Impact factor: 5.006

8.  CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil.

Authors:  Helena Almqvist; Hanna Axelsson; Rozbeh Jafari; Chen Dan; André Mateus; Martin Haraldsson; Andreas Larsson; Daniel Martinez Molina; Per Artursson; Thomas Lundbäck; Pär Nordlund
Journal:  Nat Commun       Date:  2016-03-24       Impact factor: 14.919

9.  Determining direct binders of the Androgen Receptor using a high-throughput Cellular Thermal Shift Assay.

Authors:  Joseph Shaw; Mathew Leveridge; Charlotta Norling; Jakob Karén; Daniel Martinez Molina; Daniel O'Neill; James E Dowling; Paul Davey; Suzanna Cowan; Michael Dabrowski; Martin Main; Davide Gianni
Journal:  Sci Rep       Date:  2018-01-09       Impact factor: 4.379

10.  Positioning High-Throughput CETSA in Early Drug Discovery through Screening against B-Raf and PARP1.

Authors:  Joseph Shaw; Ian Dale; Paul Hemsley; Lindsey Leach; Nancy Dekki; Jonathan P Orme; Verity Talbot; Ana J Narvaez; Michal Bista; Daniel Martinez Molina; Michael Dabrowski; Martin J Main; Davide Gianni
Journal:  SLAS Discov       Date:  2018-12-13       Impact factor: 3.341
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  1 in total

Review 1.  Biophysical Approaches for the Characterization of Protein-Metabolite Interactions.

Authors:  Anja Thalhammer; Nina K Bröker
Journal:  Methods Mol Biol       Date:  2023
  1 in total

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