| 00:00:00 | Use of isothermal titration calorimetry to investigate and identify novel peptide substrates for prolyl carboxypeptidase: a technology comparison |
| 00:01:23 | Use of isothermal titration calorimetry to investigate and identify novel peptide substrates for propyl carboxypeptidase. |
| 00:01:40 | Introduction |
| 00:02:00 | Introduction |
| 00:02:13 | Introduction |
| 00:02:17 | Introduction |
| 00:02:27 | Introduction |
| 00:02:39 | Introduction |
| 00:02:57 | Introduction |
| 00:03:08 | Aims |
| 00:03:14 | Aims |
| 00:03:17 | Aims |
| 00:03:22 | Aims |
| 00:03:26 | Aims |
| 00:03:42 | Aims |
| 00:04:05 | Aims |
| 00:04:17 | Isothermal Titration Calorimetry (ITC) |
| 00:04:38 | Isothermal Titration Calorimetry (ITC) |
| 00:04:45 | Isothermal Titration Calorimetry (ITC) |
| 00:04:54 | Isothermal Titration Calorimetry (ITC) |
| 00:05:02 | Isothermal Titration Calorimetry (ITC) |
| 00:05:31 | Isothermal Titration Calorimetry (ITC) |
| 00:05:37 | Isothermal Titration Calorimetry (ITC) |
| 00:05:41 | Isothermal Titration Calorimetry (ITC) |
| 00:05:44 | Isothermal Titration Calorimetry (ITC) |
| 00:05:53 | Isothermal Titration Calorimetry (ITC) |
| 00:06:15 | Isothermal Titration Calorimetry (ITC) |
| 00:06:20 | Isothermal Titration Calorimetry (ITC) |
| 00:06:23 | Isothermal Titration Calorimetry (ITC) |
| 00:06:26 | Isothermal Titration Calorimetry (ITC) |
| 00:06:32 | Isothermal Titration Calorimetry (ITC) |
| 00:06:36 | Isothermal Titration Calorimetry (ITC) |
| 00:06:42 | Isothermal Titration Calorimetry (ITC) |
| 00:06:50 | Isothermal Titration Calorimetry (ITC) |
| 00:07:05 | Isothermal Titration Calorimetry (ITC) |
| 00:07:31 | Isothermal Titration Calorimetry (ITC) |
| 00:07:38 | Isothermal Titration Calorimetry (ITC) |
| 00:07:51 | Isothermal Titration Calorimetry (ITC) |
| 00:07:55 | Isothermal Titration Calorimetry (ITC) |
| 00:08:00 | Isothermal Titration Calorimetry (ITC) |
| 00:08:15 | 1. Determine peptide and enzyme concentrations |
| 00:08:32 | 1. Determine peptide and enzyme concentrations |
| 00:08:38 | 1. Determine peptide and enzyme concentrations |
| 00:08:57 | 1. Determine peptide and enzyme concentrations |
| 00:09:05 | 1. Determine peptide and enzyme concentrations |
| 00:09:11 | 1. Determine peptide and enzyme concentrations |
| 00:09:18 | 1. Determine peptide and enzyme concentrations |
| 00:09:34 | 1. Determine peptide and enzyme concentrations |
| 00:09:38 | 1. Determine peptide and enzyme concentrations |
| 00:09:41 | 1. Determine peptide and enzyme concentrations |
| 00:09:46 | 1. Determine peptide and enzyme concentrations |
| 00:09:51 | 1. Determine peptide and enzyme concentrations |
| 00:09:58 | 1. Determine peptide and enzyme concentrations |
| 00:10:02 | 1. Determine peptide and enzyme concentrations |
| 00:10:16 | 1. Determine peptide and enzyme concentrations |
| 00:10:27 | 2. Determine total molar enthalpy |
| 00:10:43 | 2. Determine total molar enthalpy |
| 00:10:53 | 2. Determine total molar enthalpy |
| 00:11:02 | 2. Determine total molar enthalpy |
| 00:11:07 | 2. Determine total molar enthalpy |
| 00:11:30 | 2. Determine total molar enthalpy |
| 00:11:36 | 2. Determine total molar enthalpy |
| 00:11:40 | 2. Determine total molar enthalpy |
| 00:11:51 | 2. Determine total molar enthalpy |
| 00:11:59 | 2. Determine total molar enthalpy |
| 00:12:05 | 2. Determine total molar enthalpy |
| 00:12:29 | 3. Perform ITC experiment |
| 00:12:39 | 3. Perform ITC experiment |
| 00:12:44 | 3. Perform ITC experiment |
| 00:12:52 | 3. Perform ITC experiment |
| 00:13:11 | Results |
| 00:13:15 | 1. Hydrolysis of angiotensin III by rPRCP |
| 00:13:22 | 1. Hydrolysis of angiotensin III by rPRCP |
| 00:13:41 | 1. Hydrolysis of angiotensin III by rPRCP |
| 00:13:51 | 1. Hydrolysis of angiotensin III by rPRCP |
| 00:14:02 | 1. Hydrolysis of angiotensin III by rPRCP |
| 00:14:16 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:14:23 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:14:30 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:14:37 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:14:48 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:14:57 | 2. Hydrolysis of (pyr)-apelin-13 by rPRCP |
| 00:15:09 | 3. Method comparison |
| 00:16:02 | 3. Method comparison |
| 00:16:30 | 3. Method comparison |
| 00:17:09 | Conclusion |
| 00:17:17 | Conclusion |
| 00:17:23 | Conclusion |
| 00:17:30 | Conclusion |
| 00:17:49 | Acknowledgements |
| 00:18:09 | Acknowledgements |
| 00:18:17 | Thank you for your attentionAny questions? |
| 00:20:11 | Untitled |
| 00:21:21 | Thank you for your attentionAny questions? |
| 00:21:46 | Contact Information |
The search for novel peptide substrates for proline-specific peptidases has been hampered by a lack of suitable assays. Existing technologies are often too expensive, labor-intensive or require derivatization and separation of the reaction products. The purpose of this study was to validate the use of isothermal titration calorimetry (ITC) to study enzyme kinetics, in comparison to more conventional methods (e.g. MALDI-TOF mass spectrometry, quantitation of released C-terminal Phe using RP-HPLC). The study led to the discovery of a novel substrate for prolyl carboxypeptidase, an enzyme involved in body weight regulation.