Binding curves by continuous gradient flow-mix calorimetry
| Autor: | Antonio Parody-Morreale, Obdulio López-Mayorga, Jose C. Martinez, Jose A. Gavira |
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| Rok vydání: | 2005 |
| Předmět: |
Chromatography
Chemistry Analytical chemistry Hydrochloric acid Calorimetry Binding Condensed Matter Physics Signal Calorimeter chemistry.chemical_compound Thermodynamics Titration Ribonuclease A Deconvolution Physical and Theoretical Chemistry Flow calorimetry Thermal analysis Constant (mathematics) Instrumentation Protein–ligand interactions |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | 1 table, 4 figures. teady-state flow-mix calorimetric methods require large amounts of sample and time to obtain data at variable reactant ratios. The use of continuous concentration gradients allows rapid generation of data with chemicals consumption comparable to other calorimetric techniques. An experimental set-up able to generate a linear concentration gradient of one reactant, while keeping the other concentration constant during the experiment, permits a complete titration in a single experiment. Since the calorimeter is not operating under steady-state conditions, the output is affected by the dynamics of the instrument. Therefore, it is necessary to correct the experimental curve by deconvolution to obtain the true signal. Titration of the carboxyl group of glycine with hydrochloric acid, as well as the binding of ribonuclease A to two different nucleotides prove the efficacy of this new methodology. We thank to M. Balmacz for revising the English text. This work was financed by grant BIO2003-04274 from the Spanish Ministry of Science and Education, and grant HPRN-CT-2002-00241 from the European Union; it was also financed, in part, by FEDER funds from the European Union. J.A.G. was supported by the Andalusian Regional government, Spain. We also acknowledge Dr. Lee Hansen for his careful and exhaustive revision of the manuscript. |
| Databáze: | OpenAIRE |
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