18 resultados para Magnetic Circular-dichroism
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Biophysical Chemistry 110 (2004) 83–92
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Journal of Biological Inorganic Chemistry (2010)15: 271-281
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Dissertation for the Master’s Degree in Structural and Functional Biochemistry
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The focus of this Thesis was the study of the sensor domains of two heme-containing methyl-accepting chemotaxis proteins (MCP) from Geobacter sulfurreducens: GSU0582 and GSU0935. These domains contain one c-type heme, form swapped dimers with a PAS-like fold and are the first examples of a new class of heme sensors. NMR spectroscopy was used to assign the heme and polypeptide signals in both sensors, as a first step to probe conformational changes in the vicinity of the hemes. However, the presence of two conformations in solution impaired the confident assignment of the polypeptide signals. To understand how conformational changes and swapped dimerization mechanism can effectively modulate the function of the two sensor domains and their signal transduction process, the sensor domains folding and stability were studied by circular dichroism and UV-visible spectroscopy. The results showed differences in the thermodynamic stability of the sensors, with GSU0582 displaying higher structural stability. These studies also demonstrated that the heme moiety undergoes conformational changes matching those occurring at the global protein structure and that the content of intrinsically disordered segments within these proteins (25% for GSU0935; 13% for GSU0582) correlates with the stability differences observed. The thermodynamic and kinetic properties of the sensor domains were determined at different pH and ionic strength by visible spectroscopy and stopped-flow techniques. Despite the remarkably similar spectroscopic and structural features of the two sensor domains, the results showed that their properties are quite distinct. Sensor domain GSU0935 displayed more negative reduction potentials and smaller reduction rate constants, which were more affected by pH and ionic strength. The available structures were used to rationalize these differences. Overall, the results described in this Thesis indicate that the two G. sulfurreducens MCP sensor domains are designed to function in different working potential ranges, allowing this bacterium to trigger an adequate cellular response in distinct anoxic subsurface environments.
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Dissertação apresentada para obtenção do grau de Doutor em Matemática na especialidade de Equações Diferenciais, pela Universidade Nova de Lisboa,Faculdade de Ciências e Tecnologia
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do Grau de Mestre em Engenharia Mecânica
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Dissertation submitted in Faculdade de Ciências e Tecnologia of Universidade Nova de Lisboa for the degree of Master in Biomedical Engineering
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Biotecnologia
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do Grau de Mestre em Engenharia Mecânica
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Dissertação para obtenção do Grau de Mestre em Biotecnologia
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Dissertation to Obtain the Degree of Master in Biomedical Engineering
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Based on the report for “Project IV” unit of the PhD programme on Technology Assessment (Doctoral Conference) at Universidade Nova de Lisboa (December 2011). This thesis research has the supervision of António Moniz (FCT-UNL and ITAS-KIT) and Michael Decker (Karlsruhe Institute of Technology-ITAS). Other members of the thesis committee are Carlos Alberto da Silva (University of Évora), José Maria de Albuquerque (Institute of Welding and Quality), Lotte Steuten (University of Twente), Mário Forjaz Secca (FCT-UNL) and Nelson Chibeles Martins (FCT-UNL).
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Based on the report for the unit “Project III” of the PhD programme on Technology Assessment in 2011. The unit was supervised by Prof. António B. Moniz (FCT-UNL).
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Biochemistry. 2008 Oct 14;47(41):10852-62. doi: 10.1021/bi801375q