Expression, purification and crystallization of class C acid phosphatases from Francisella tularensis and Pasteurella multocida
| Autor: | Harkewal Singh, Michael J. Calcutt, John J. Tanner, Thomas J. Reilly, Richard L. Felts, Li Ma, Thomas J. Malinski |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Pasteurella multocida
Phosphatase Acid Phosphatase Biophysics Crystallography X-Ray Biochemistry Microbiology Structural Biology Genetics Francisella tularensis Pathogen chemistry.chemical_classification Cloning biology Acid phosphatase Condensed Matter Physics biology.organism_classification Enzyme chemistry Cytoplasm Crystallization Communications biology.protein Crystallization |
| Popis: | Class C nonspecific acid phosphatases are bacterial enzymes that are secreted across the cytoplasmic membrane and hydrolyze a variety of phosphomonoesters at acidic pH. These enzymes are of interest for the development of improved vaccines and clinical diagnostic methods. In one case, the category A pathogen Francisella tularensis, the class C phosphatase plays a role in bacterial fitness. Here, the cloning, expression, purification and crystallization methods for the class C acid phosphatases from F. tularensis and Pasteurella multocida are reported. Crystals of the F. tularensis enzyme diffracted to 2.0 A resolution and belonged to space group C222(1), with one enzyme molecule in the asymmetric unit. Crystals of the P. multocida enzyme diffracted to 1.85 A resolution and belonged to space group C2, with three molecules in the asymmetric unit. Diffraction patterns from crystals of the P. multocida enzyme exhibited multiple interpenetrating reciprocal-space lattices, indicating epitaxial twinning. Despite this aberrance, autoindexing was robust and the data could be satisfactorily processed to 1.85 A resolution using MOSFLM and SCALA. |
| Databáze: | OpenAIRE |
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