Stability and structural characterization of the FlgD protein from Helicobacter pylori

Stojanović, Mario (2016) Stability and structural characterization of the FlgD protein from Helicobacter pylori. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

FlgD protein from Helicobacter pylori (HpFlgD) is a chaperon, which plays a role during hook assembly. The hook is a part of the motility apparatus and is also a virulence factor. The lack of the flgD gene makes bacteria immotile and that fact highlights the importance of the FlgD protein during synthesis of flagella. This thesis describes the transformation of Escherichia coli with the vector with sequence coding for HpFlgD His6, over-expresion of protein, isolation by affinity chromatography and gel-filtration methods, and crystallization trials with a pure protein sample. During the crystallization trials the following conditions were varied: concentration of PEG 1500 (w/v), pH of the SPG buffer, temperature (4 °C and 16 °C), and concentration of the protein. The data obtained by the X-ray structure analysis of FlgD protein enabled us to make a structural model up to 2,86 Å resolution. The solved structure of the HpFlgD protein is similar to the two already known structures of FlgD proteins from Xanthomonas campestris and Pseudomonas aeruginosa. The protein has two domains, Tudor and Fn III, which are built up of β sheets and loops. The main difference between the two known structures and this one is the rotated position of the Tudor domain.

Item Type: Thesis (Diploma thesis)
Keywords: human pathogen, protein purification, protein crystallization, X-ray diffraction, protein structure
Supervisor: Matković Čalogović, Dubravka
Co-supervisor: Kekez, Ivana
Date: 2016
Number of Pages: 50
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 24 Mar 2016 15:09
Last Modified: 24 Mar 2016 15:09
URI: http://digre.pmf.unizg.hr/id/eprint/4623

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