Competitive DNA binding of Ru(bpy)2dppz2+ enantiomers studied with isothermal titration calorimetry (ITC) using a direct and general binding isotherm algorithm

A simple algorithm allowing for binding isotherm calculations of almost any level of complexity is demonstrated here in a competitive ITC setting with enantiopure Ru-bpy intercalating into AT-DNA.

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Insights into the Relative DNA Binding Affinity and Preferred Binding Mode of Homologous Compounds Using Isothermal Titration Calorimetry (ITC)

Applications of Calorimetry in a Wide Context - Differential Scanning Calorimetry, Isothermal Titration Calorimetry and Microcalorimetry ◽

10.5772/54061 ◽

2013 ◽

Cited By ~ 1

Author(s):

Ruel E.

Keyword(s):

Dna Binding ◽

Binding Affinity ◽

Isothermal Titration Calorimetry ◽

Binding Mode ◽

Titration Calorimetry ◽

Homologous Compounds ◽

Dna Binding Affinity

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Specific DNA binding by the homeodomain Nkx2.5(C56S): Detection of impaired DNA or unfolded protein by isothermal titration calorimetry

Proteins Structure Function and Bioinformatics ◽

10.1002/prot.20960 ◽

2006 ◽

Vol 64 (1) ◽

pp. 13-18 ◽

Cited By ~ 2

Author(s):

Elfrieda Fodor ◽

Ann Ginsburg

Keyword(s):

Dna Binding ◽

Isothermal Titration Calorimetry ◽

Titration Calorimetry ◽

Unfolded Protein

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Recommendations on measurement and analysis of results obtained on biological substances using isothermal titration calorimetry (IUPAC Technical Report)

Pure and Applied Chemistry ◽

10.1351/pac200880092025 ◽

2008 ◽

Vol 80 (9) ◽

pp. 2025-2040 ◽

Cited By ~ 21

Author(s):

Frederick P. Schwarz ◽

Timm Reinisch ◽

Hans-Jürgen Hinz ◽

Avadhesha Surolia

Keyword(s):

Isothermal Titration Calorimetry ◽

Small Molecule ◽

Binding Constants ◽

Titration Calorimetry ◽

Heat Of Mixing ◽

Buffer Solution ◽

Binding Model ◽

Binding Isotherm ◽

Analyte Solution ◽

Sample Vessel

Isothermal titration calorimetry (ITC) is widely used to determine the thermodynamics of biological interactions including protein-protein, small molecule-protein, protein-DNA, small molecule-DNA, and antigen-antibody interactions. An ITC measurement consists of monitoring the transfer of heat between an analyte solution in a sample vessel and a reference solution in a reference vessel upon injection of a small aliquot of titrant solution into the sample vessel at a fixed ITC operating temperature. A binding isotherm is generated from the heat-transferred-per-injection data and values for the binding constants, the apparent binding enthalpies, and the apparent ratio of the amount of titrant to analyte for the binding reaction are then determined from fits of a binding model, whether it is a single site, identical multi-site, or an interacting multi-site binding model, to the binding isotherm. Prior to the fitting procedure, corrections should be made for contributions from extraneous heat of mixing determined separately from injections of the titrant into just the dialysate/buffer solution. Ultra-high binding constants, which cannot be directly determined from an ITC measurement, can be determined by a displacement ITC method where injections of the tight-binding titrant into a solution of a weaker-binding titrant-analyte complex displaces the weaker-binding titrant from the complex. The Michaelis and catalytic constants can be determined for an enzyme reaction from injections of a substrate or enzyme titrant into an enzyme or substrate analyte solution. Several binding reactions are suggested to check the operating performance of the ITC. The reporting of ITC results must be specific with regard to the composition of the titrant and the analyte solutions, the temperature, and the model used in the analysis.

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