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X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
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X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
Capítulos
- 00:04Introduction
- 01:03Activity Assay
- 02:19Apoenzyme Preparation
- 03:32Dimer Preparation
- 04:03Microseeding
- 05:01X-Ray Diffraction
- 05:48Indexation
- 06:50Molecular Replacement
- 08:04Model Building
- 09:07Results: Representative Thermotoga Maritima M42 Aminopeptidase TmPep1050 Oligomeric State Transition
- 10:50Conclusion
This protocol has been developed to study the dimer-dodecamer transition of TmPep1050, an M42 aminopeptidase, at the structural level. It is a straightforward pipeline starting from protein purification to X-ray data processing. Crystallogenesis, data set indexation, and molecular replacement have been emphasized through a case of study, TmPep1050H60A H307A variant.
