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Thermodynamics and solvent linkage of macromolecule-ligand interactions.
Duff, Michael R; Howell, Elizabeth E.
Afiliación
  • Duff MR; Department of Biochemistry, Cellular, and Molecular Biology, University of Tennessee, Knoxville, TN 37996-0840, United States.
  • Howell EE; Department of Biochemistry, Cellular, and Molecular Biology, University of Tennessee, Knoxville, TN 37996-0840, United States. Electronic address: lzh@utk.edu.
Methods ; 76: 51-60, 2015 Apr.
Article en En | MEDLINE | ID: mdl-25462561
Binding involves two steps, desolvation and association. While water is ubiquitous and occurs at high concentration, it is typically ignored. In vitro experiments typically use infinite dilution conditions, while in vivo, the concentration of water is decreased due to the presence of high concentrations of molecules in the cellular milieu. This review discusses isothermal titration calorimetry approaches that address the role of water in binding. For example, use of D2O allows the contribution of solvent reorganization to the enthalpy component to be assessed. Further, the addition of osmolytes will decrease the water activity of a solution and allow effects on Ka to be determined. In most cases, binding becomes tighter in the presence of osmolytes as the desolvation penalty associated with binding is minimized. In other cases, the osmolytes prefer to interact with the ligand or protein, and if their removal is more difficult than shedding water, then binding can be weakened. These complicating layers can be discerned by different slopes in ln(Ka) vs osmolality plots and by differential scanning calorimetry in the presence of the osmolyte.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Solventes / Agua / Colorimetría Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2015 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Solventes / Agua / Colorimetría Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2015 Tipo del documento: Article País de afiliación: Estados Unidos