Influence of dynamic power compensation in an isothermal titration microcalorimeter.
A theoretical analysis in Laplace's transformed domain based on a power balance represents a suitable model for an isothermal titration calorimeter with dynamic power compensation, designed and implemented in our laboratory. A rigorous calibration of the injection system and the calorimetric response was also made. Using electrically generated heat pulses, two different time constants have been determined from the calorimetric transfer function and assigned to the physical parts of the calorimeter. The same was done for a protein-ligand interaction. The binding of 2'-CMP to ribonuclease A at low and high ionic strengths was used to check the apparatus and the results were compared with those obtained by other authors (Wiseman, T.; Williston, S.; Brandts, J.F.; Lung-Nan, L. Anal. Biochem. 1989, 179, 131-137). In this case, the analysis showed a different time constant for the heat source. Independently of the nature of the heat source, the calorimetric time constants obtained while working under compensation are always smaller than those corresponding to a noncompensated system. The improvement of the calorimetric response introduced by dynamic power compensation is thus explained in terms of the reduction of the time constants characteristic of the calorimeter. This theoretical model can be used to predict the shape of the thermogram for any given reaction of either known or supposed thermodynamic parameters. Therefore, the calorimetric study is extended to the other nucleotides, 2'-UMP and 5'-dUMP, which have not hitherto been reported in the literature.
García-Fuentes
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L
Departamento de Química Física, Bioquímica y Química Inorgánica, Facultad de Ciencias Experimentales, Universidad de Almería, Spain. lgarcia@ualm.es
Barón
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C
Mayorga
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OL
D002138:Calibration; D002151:Calorimetry / Q000295:instrumentation*; D003568:Cytidine Monophosphate / Q000032:analysis / Q000737:chemistry*; D003856:Deoxyuracil Nucleotides / Q000737:chemistry; D008962:Models, Theoretical; D009994:Osmolar Concentration; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D017186:Titrimetry / Q000379:methods; D014542:Uridine Monophosphate / Q000737:chemistry
D003856:Deoxyuracil Nucleotides; C007267:2'-deoxyuridylic acid; D014542:Uridine Monophosphate; D012259:Ribonuclease, Pancreatic; D003568:Cytidine Monophosphate