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Title: Protein–ligand interactions investigated by thermal shift assays (TSA) and dual polarization interferometry (DPI)

The biophysical characterization of protein–ligand interactions in solution using techniques such as thermal shift assay, or on surfaces using, for example, dual polarization interferometry, plays an increasingly important role in complementing crystal structure determinations. Over the last decades, a wide range of biophysical techniques investigating protein–ligand interactions have become indispensable tools to complement high-resolution crystal structure determinations. Current approaches in solution range from high-throughput-capable methods such as thermal shift assays (TSA) to highly accurate techniques including microscale thermophoresis (MST) and isothermal titration calorimetry (ITC) that can provide a full thermodynamic description of binding events. Surface-based methods such as surface plasmon resonance (SPR) and dual polarization interferometry (DPI) allow real-time measurements and can provide kinetic parameters as well as binding constants. DPI provides additional spatial information about the binding event. Here, an account is presented of new developments and recent applications of TSA and DPI connected to crystallography.
Authors:
;  [1] ;  [2] ;  [1]
  1. Durham University, South Road, Durham DH1 3LE (United Kingdom)
  2. Biolin Scientific, 62 Wellington Road South, Stockport, Cheshire SK1 3SU (United Kingdom)
Publication Date:
OSTI Identifier:
22347736
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 1; Other Information: PMCID: PMC4304684; PMID: 25615858; PUBLISHER-ID: ba5217; OAI: oai:pubmedcentral.nih.gov:4304684; Copyright (c) Grøftehauge et al. 2015; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CALORIMETRY; CRYSTAL STRUCTURE; CRYSTALLOGRAPHY; CRYSTALS; CURRENTS; INTERACTIONS; INTERFEROMETRY; LIGANDS; MATHEMATICAL SOLUTIONS; POLARIZATION; RESOLUTION; RESONANCE; SURFACES; TIME MEASUREMENT