Label-Free Determination of the Dissociation Constant of Small Molecule-Aptamer Interaction by Isothermal Titration Calorimetry

2016 ◽  
pp. 113-125 ◽  
Author(s):  
Marc Vogel ◽  
Beatrix Suess
Keyword(s):  
Dissociation Constant ◽  
Isothermal Titration Calorimetry ◽  
Small Molecule ◽  
Titration Calorimetry ◽  
Label Free

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

2008 ◽  
Vol 80 (9) ◽  
pp. 2025-2040 ◽  
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.

Enzymatic activity assay of d-hydantoinase by isothermal titration calorimetry. Determination of the thermodynamic activation parameters for the hydrolysis of several substrates

2006 ◽  
Vol 67 (1) ◽  
pp. 57-66 ◽  
Author(s):  
Montserrat Andújar-Sánchez ◽  
Francisco Javier Las Heras-Vázquez ◽  
Josefa María Clemente-Jiménez ◽  
Sergio Martínez-Rodríguez ◽  
Ana Camara-Artigas ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Isothermal Titration Calorimetry ◽  
Activation Parameters ◽  
Titration Calorimetry ◽  
Activity Assay ◽  
Thermodynamic Activation Parameters ◽  
Hydrolysis Of ◽  
Enzymatic Activity Assay

1P266 Measurement of anion binding dissociation constant of pharaonis halorhodopsin using isothermal titration calorimetry

2005 ◽  
Vol 45 (supplement) ◽  
pp. S98
Author(s):  
S. Hayashi ◽  
S. Miyauchi ◽  
C. Hasegawa ◽  
M. Demura ◽  
N. Kamo
Keyword(s):  
Dissociation Constant ◽  
Isothermal Titration Calorimetry ◽  
Anion Binding ◽  
Titration Calorimetry

Isothermal titration calorimetry determination of individual rate constants of trypsin catalytic activity

2015 ◽  
Vol 479 ◽  
pp. 18-27 ◽  
Author(s):  
César Aguirre ◽  
Itzel Condado-Morales ◽  
Luis F. Olguin ◽  
Miguel Costas
Keyword(s):  
Catalytic Activity ◽  
Isothermal Titration Calorimetry ◽  
Rate Constants ◽  
Titration Calorimetry ◽  
Individual Rate
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