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dc.contributor.authorLevin, Benjamin Diamonen_US
dc.date.accessioned2016-01-26T18:57:19Z
dc.date.available2016-01-26T18:57:19Z
dc.date.issued2013
dc.identifier.urihttps://hdl.handle.net/2144/14093
dc.description.abstractHeme containing proteins and their reactivity play a central role in biological systems; they have a vast range of functions including electron transfer, catalysis, and respiration. Cytochromes c and heme c containing proteins have been used widely as model systems to understand how structure and dynamics lead toward function. In this thesis, a variety of biophysical methods are used to investigate two heme c containing model systems to gain insight into how redox potential and reactivity are modulated through changes in the local environment. Mitochondrial cytochrome c undergoes several pH dependent conformational rearrangements that involve different heme ligation and have associated changes in redox potential. Under basic conditions (pH greater than 8), the axial methionine (Met) residue is replaced by one of several nitrogen based ligands, usually a nearby lysine residue, and is coined the "alkaline transition". It is accompanied by a large downward shift in redox potential. The functional utility of this conformational change is not fully understood however it is strongly implicated in the signaling cascade for apoptosis. Bacterial monoheme cytochromes c exhibit similar phenomenological Met-loss behavior as a function of electrode material. In Chapter 2 we utilize Hydrogenobacter thermophilus cytochrome c552 as a model system for the assessment of redox thermodynamics and changes in redox potential associated with the Met-loss form. In Chapter 3 we extend our investigation to homologous cytochromes c. Bacterial cytochrome c peroxidases catalyze the two-electron reduction of hydrogen peroxide to water utilizing cytochrome c as an endogenous electron donor. Chapter 4 describes the first recombinant construct of the diheme Nitrosomonas europaea cytochrome c peroxidase (Ne CCP); a defining family member of constitutively active cytochrome c peroxidases. A variety of biophysical techniques were used to confirm similarity between the recombinant Ne CCP and native enzyme. Chapter 5 extends our investigation to the role of constitutively conserved glutamine and glutamic acid residues within the active site, and two conserved tryptophan residues; the first situated between hemes and the second distal to the active site. In Chapter 6, stopped flow spectroscopy is used to investigate the first intermediates of the Ne CCP catalytic mechanism.en_US
dc.language.isoen_US
dc.subjectChemistryen_US
dc.subjectCyclic voltammetryen_US
dc.subjectCytochrome cen_US
dc.subjectDiheme peroxidaseen_US
dc.subjectProtein film voltammetryen_US
dc.titleProbing the structure-function relationship of heme c containing bacterial proteins: monoheme cytochromes c and diheme cytochrome c peroxidaseen_US
dc.typeThesis/Dissertationen_US
dc.date.updated2016-01-22T18:53:43Z
etd.degree.nameDoctor of Philosophyen_US
etd.degree.leveldoctoralen_US
etd.degree.disciplineChemistryen_US
etd.degree.grantorBoston Universityen_US


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