The aim of the thesis was to compare the haloalkane dehalogenase DhaA106 with other studied protein mutant variants. For diffraction analysis, it was necessary to growappropriate crystals of DhaA106 protein. Crystallization experiments were performed by standard sitting drop method. Obtained rod crystals were used for diffraction analysis. During diffraction measurement completeset of 500 diffraction images were obtained, from which the electron density map was formed and the spatial model of the molecular structure of DhaA106 dehalogenase was created. Alternative conformation of amino acids, water molecules, parts of precipitating solution and chloride ion inthe active sitewere subsequently added into the model structure. The final structure of the moleculewas refined and validated. Subsequently, the amino acids pentade in the active site and whole protein structure were analyzed . Values of validation and refining and interatomic distances in the active site were compared with several variants mutant dehalogenase DhaA, as well as the precipitating solutions used for crystallization and diffraction data collection. In the last step of the work interactions of the selected substrate with the protein surface in the vicinity of the tunnel connecting the active site of the surfacewere studied. Mentioned interaction study was based on the principle of combination of the Monte-Carlo methods with protein structure prediction algorithms.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:188603 |
| Date | January 2015 |
| Creators | MALCHER, Pavel |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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