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Biochemistry

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JoVE Journal Biochemistry
Production, Crystallization and Structure Determination of C. difficile PPEP-1 via Microseeding and Zinc-SAD
 

Production, Crystallization and Structure Determination of C. difficile PPEP-1 via Microseeding and Zinc-SAD

Article DOI: 10.3791/55022-v • 13:34 min • December 30th, 2016
December 30th, 2016 •
1, 1, 1, 1
1Institute of Biochemistry, University of Cologne

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Proline-proline endopeptidase-1 (PPEP-1) is a secreted metalloprotease and promising drug-target from the human pathogen Clostridium difficile. Here we describe all methods necessary for the production and structure determination of this protein.

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Production Crystallization Structure Determination C. Difficile PPEP-1 Microseeding Zinc-SAD Recombinant Proline Proline Endopeptidase-1 RPPEP-1 Lattice Diffraction Quality Crystals X-ray Diffraction Analysis Microcity Method Substrate Peptide Complexes Cross Seeding Experiments Protein Variance Cocrystallization Experiments Handling Of Crystallization Methods Sitting Drop Format Commercially Available Screens Crystallization Robot Centrifugal Ultrafiltration Device
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