By Dr. John J. Correia and Dr. H. William Detrich, III (Eds.)

ISBN-10: 0123725208

ISBN-13: 9780123725202

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Additional resources for Biophysical Tools for Biologists, Volume One: In Vitro Techniques

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To achieve this fraction of unbound ligand, it is necessary to work at ligand concentrations approaching the reciprocal of the binding constant and quickly becomes impractical for ultratight binding interactions. DSC requires neither a ligand signal nor any fraction of unbound ligand. The methods discussed above were applied to the study of protein–ligand interactions but are, in fact, quite general in scope and can easily be applied to both protein and DNA systems with only subtle changes in the models to account, for example, for the lattice 12 Nichola C.

Measurement of small ligand binding to a regulatory protein provides information about the binding mechanism and energetics of the binding process. Mechanism refers to the number of ligands that bind and whether, for multiple ligand binding, any cooperativity exists. Knowledge of the binding energetics allows one to relate the probability of occupancy of the protein by ligand to physiological concentrations of both the regulatory protein and small ligand. There are many methods for measuring small ligand binding which range from the low-tech equilibrium dialysis method to more sophisticated methods such as isothermal titration calorimetry (ITC).

Methods Enzymol. 340, 193–211. Turnbull, W. , and Daranas, A. H. (2003). On the value of c: Can low aYnity systems be studied by isothermal titration calorimetry? J. Am. Chem. Soc. 125, 14859–14866. , and Freire, E. (2001). The binding energetics of first- and secondgeneration HIV-1 protease inhibitors: Implications for drug design. Arch. Biochem. Biophys. 390, 169–175. , Todd, M. , and Freire, E. (2000). HIV-1 protease inhibitors: Enthalpic versus entropic optimization of the binding aYnity. Biochemistry 39, 2201–2207.

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Biophysical Tools for Biologists, Volume One: In Vitro Techniques by Dr. John J. Correia and Dr. H. William Detrich, III (Eds.)


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