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JoVE Journal Biochemistry
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

Article DOI: 10.3791/61288-v • 11:27 min • May 13th, 2020
May 13th, 2020 •
1,2, 2, 1, 1
1Laboratory of Microbiology, Department of Molecular Biology, Université Libre de Bruxelles, 2Labiris Institut de Recherche

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Summary
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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.

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X-Ray Crystallography Oligomeric State Transition Thermotoga Maritima M42 Aminopeptidase TmPep1050 Aminopeptidases Oligomeric States Oligomerization-dependent Activities Inactive Dimers Active Dodecamers Conventional Technique Beamline X-ray Crystallography Facility Special Devices Protein Degree Of Oligomerization X-ray Data Processing Crystallography Understanding Trainings XDS Wiki Phenix Tutorials Activity Assay L-Leucine P-Nitroanilide MOPS Cobalt Two Chloride Methanol Incubation Temperature Acid Stop Reaction
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