title: Biochemical and Biophysical Characterization of the Human Ethylmalonic Encephalopathy non-Heme Sulfur [Fe]-Dioxygenase ETHE1, and X-ray Absorption Spectroscopy Applications and Methods Development
creator: Salomone-Stagni, Marco
subject: 570
subject: 570 Life sciences
description: This work focuses on the biochemical and biophysical characterization of highly interesting metallo-proteins and is divided in two main projects. The first project regards the characterization of the iron-based enzyme ETHE1 from Homo sapiens, which is fundamental for detoxification from highly reactive persulfide species and whose specific mutations on the structural gene or on its regulatory sequences cause ethylmalonic encephalopathy. Moreover, ETHE1 was reported to be involved in apoptosis and cancer through the interaction with transcription factors such as NF-kB and p53. The protein has been studied by a number of techniques, noticeably XAS (Xray-absorption spectroscopy), SAXS (Small Angle X-ray Scattering Spectroscopy), Mössbauer spectroscopy and ITC (Iso-Thermal Calorimetry). The second project is based on XAS analysis and method development applied to metallo-proteins that are likely to have a substantial impact on basic and applied science. The mononuclear [Fe]-hydrogenase Hmd from Methanocaldococcus jannaschii has been extensively investigated by XANES and EXAFS analysis in the native and inhibited forms. This study revealed the major determiners of the electronic structure of this unique hydrogenase iron site and shed light on the chemicophysical requirements for the heterolysis of molecular hydrogen. In addition, I contribute to the characterization of the optically excited state of a bis (μ-oxo)-dicopper(III) complex mimicking the potential di-oxo to μ-oxo transition in tyrosinases active site. We took advantage of two cooperative approaches: pumped-XAS and an innovative combination of EXAFS spectroscopy and resonant Raman scattering. I present also the XAS analysis results obtained on the protein ABCE1, which is one of the most conserved proteins in the evolution of eukaryotes and archaea. ABCE1 is essential for cell viability and is a unique ATP-binding-cassette protein bearing iron-sulfur clusters.
date: 2010
type: Dissertation
type: info:eu-repo/semantics/doctoralThesis
type: NonPeerReviewed
format: application/pdf
identifier: https://archiv.ub.uni-heidelberg.de/volltextserverhttps://archiv.ub.uni-heidelberg.de/volltextserver/11243/1/Marco_Salomone_Stagni_PhD_Thesis.pdf
identifier: DOI:10.11588/heidok.00011243
identifier: urn:nbn:de:bsz:16-opus-112436
identifier:   Salomone-Stagni, Marco  (2010) Biochemical and Biophysical Characterization of the Human Ethylmalonic Encephalopathy non-Heme Sulfur [Fe]-Dioxygenase ETHE1, and X-ray Absorption Spectroscopy Applications and Methods Development.  [Dissertation]     
relation: https://archiv.ub.uni-heidelberg.de/volltextserver/11243/
rights: info:eu-repo/semantics/openAccess
rights: http://archiv.ub.uni-heidelberg.de/volltextserver/help/license_urhg.html
language: eng