| 00:00:00 | Welcome |
| 00:01:23 | Calorimetry- Basics |
| 00:03:11 | Isothermal Titration Calorimeter |
| 00:04:55 | Enzyme Kinetics |
| 00:05:52 | Enzyme Kinetics |
| 00:07:41 | Enzyme Kinetics and ITC |
| 00:09:13 | Determination of Enthalpy- M1 |
| 00:10:12 | Experimental Considerations- M1 |
| 00:11:01 | Calculating "True" Reaction Enthalpy |
| 00:12:31 | Determining dQ/dt |
| 00:13:41 | Experimental Considerations- M2 |
| 00:14:45 | Determining dQ/dt |
| 00:15:07 | Continuous assay |
| 00:15:40 | Continuous assay |
| 00:16:42 | Experimental Considerations- Single Injection Method |
| 00:18:01 | Practical Considerations |
| 00:19:40 | Ni(ll)- Dependent Urease |
| 00:20:41 | Ni(ll)- Dependent Urease |
| 00:21:10 | Ni(ll)- Dependent Urease |
| 00:22:12 | Lactate Dehydrogenase |
| 00:22:59 | Lactate Dehydrogenase |
| 00:23:22 | Laccase |
| 00:24:30 | Enzyme Inhibition by ITC |
| 00:26:05 | Enzyme Inhibition by ITC |
| 00:26:27 | Urease Inhibition by Fluoride |
| 00:28:23 | LDH-A Inhibition by Oxamate |
| 00:29:31 | Trypsin Inhibition by Benzamidine |
| 00:30:14 | Conclusions |
| 00:31:00 | Acknowledgments |
| 00:31:16 | Thank you for your attentionAny questions? |
| 00:45:48 | Contact Information |
Isothermal Titration Calorimetry
(ITC) is a technique that measures the heat released or absorbed during a chemical reaction as an intrinsic probe to characterize any chemical process. In the present webinar, the experimental ITC-based methods to quantify kinetics and thermodynamics of enzymatic reactions are described.
ITC can concomitantly determine kinetics and thermodynamic parameters (kcat, KM, DH) of enzymatic reactions, not requiring any modification or labelling of the system under analysis, as heat changes spontaneously during enzymatic catalysis.
ITC can be performed in solution and needs little amount of material. These properties make this technique an invaluable, powerful and unique tool to extend the study of enzyme kinetics to several applications, such as drug discovery.
ITC can concomitantly determine kinetics and thermodynamic parameters (kcat, KM, DH) of enzymatic reactions, not requiring any modification or labelling of the system under analysis, as heat changes spontaneously during enzymatic catalysis.
ITC can be performed in solution and needs little amount of material. These properties make this technique an invaluable, powerful and unique tool to extend the study of enzyme kinetics to several applications, such as drug discovery.