Offerte TESI PER I CORSI DI LAUREA MAGISTRALE

The function of mitochondria is central in energy metabolism and most aspect of aerobic cell biology. The apoptosis inducing factor (AIF) is a highly  ...

The function of mitochondria is central in energy metabolism and most aspect of aerobic cell biology. The apoptosis inducing factor (AIF) is a highly conserved eukaryotic mitochondrial protein that, in addition to be involved in caspase-independent apoptosis, possesses a key role in the biogenesis and maintenance of mitochondria in healthy cells. To fulfill the latter function, AIF steadily interacts with the protein partner CHCHD4 within the intermembrane space of the organelle, where they play a chaperon-type action towards cytoplasm-imported polypeptides. In particular, the AIF-CHCHD4 complex catalyzes the redox dependent formation of disulfide bridges in the target protein, most of which are respiratory complex subunits, by specifically oxidizing some of their Cys side chains. We found that CHCHD4 dramatically affects the structure and reactivity of AIF. Aim of the present thesis project is to ascertain how CHCHD4 binding allosterically modulates the affinity of AIF for its other ligands, including NAD+/H and its redox catalytic activity. The strategy of the study will include the production of the two mammalian proteins in recombinant form, their purification by high performance chromatography, and the in vitro study of their interaction by several different biochemical techniques, including spectrophotometry, analytical chromatography, thermal shift analysis, micro scale thermophoresis, protein crystallography.