Description:
The study of structure-function relationship in proteins is considered one of the building blocks in biochemistry with important consequences in diverse fields as molecular biology, genetics, protein engineering and represents a challenge that requires a lot of effort to succeed. So far, the understanding of structure-function relationship of a newly identified pharmacological target combined both with cell-based assays and with the data collected when mutations or improper folding of such target are associated with serious diseases, are all information that are critical for drug discovery and medicine. A peculiar role in predicting the protein function of a target and for deep understanding of the molecular mechanisms in a living cell is assigned to the protein-protein interactions (PPI). Our studies are focused on the study of PPI of proteins correlated with the insurgence of pathologies taking advantage from different techniques such as expression of recombinant proteins in Escherichia coli or in mammalian or insect cells, spectrofluorimetry, circular dichroism, Elisa assays, thermophoresis, ITC, SPR and lastly NMR spectroscopy as a powerful approach for the structural characterization and for binding experiments with DNA, potential interactors or different molecules of organic or peptidic nature. In such context, among different analysed proteins, we focused on the study of the transcription factor PHOX2B whose poly-alanine triplet expansions in the coding region cause the 90% insurgence of Congenital Central Hypoventilation Syndrome (CCHS) so rendering this protein an intriguing target to understand the insurgence of this syndrome and for the design of a novel therapeutical approach. Unexpectedly, we identified the formation of fibrils only for the pathological variant, suggesting a plausible role of such fibrils in the insurgence of CCHS. Another aim of this research line is to delineate by means of molecular biology techniques, circular dichroism, ITC methodology and NMR spectroscopy the structural and functional properties of selected Intrinsically Disordered Proteins (IDPs) or Intrinsically Disordered Domains (IDDs).