- Role: Ricercatore
- Office:U3 stanza 4009
- Telephone number:02- 64483437
- Reception Hours:mer dalle 15 alle 17
2) PROTEIN STRUCTURE, STABILITY AND AGGREGATION STUDIED BY FT-IR SPECTROSCOPY
3) PROTEOMICS AND BIOPOLYMER MASS SPECTROMETRY
4) STRUCTURE, PHYSIOLOGICAL ROLE AND PATHOGENIC POTENTIAL OF AMYLOIDOGENIC PROTEINS
PROTEIN ENGINEERING AND INDUSTRIAL ENZYMOLOGY
Enzymes employed in biocatalysis, in particular lipases, and model proteins such as lactoglobulins are studied by a combined approach of mutagenesis (both directed evolution and random mutagenesis), heterologous expression, biochemical and biophysical characterization, in order to highlight the molecular bases of their stability, function and propensity to aggregate upon expression in bacterial cells. Cold adapted enzymes are used as models to understand the structural determinants of activity at low temperature. Moreover, novel biocatalysts are isolated from non commercial sources or produced by protein engineering.
Partecipants: Brocca Stefania , Lotti Marina
PROTEIN STRUCTURE, STABILITY AND AGGREGATION STUDIED BY FT-IR SPECTROSCOPY
Infrared absorption spectroscopy is used to obtain structural information on proteins in different environments, in aqueous and non aqueous solution, in solid form of pharmaceutical formulations, and within intact bacterial cells. In particular, FT-IR spectroscopy is employed to characterize the secondary structure of recombinant proteins , the stability of their different secondary elements during thermal unfolding, and the extent of glycosylation. Protein aggregation in form of inclusion bodies (IBs) in recombinant bacteria is also studied to monitor the kinetics of aggregation in vivo and to characterize the IB structural properties. Model peptides for amyloid proteins involved in neurodegenerative diseases are also investigated to identify intermediate species in the fibril formation process and to characterize at molecular level the fibril structural properties.
Partecipants: Doglia Silvia Maria
PROTEOMICS AND BIOPOLYMER MASS SPECTROMETRY
Mass spectrometry is employed on one side as an analytical tool for proteomics. The focus is on intracellular interactions and phosphorylation of the inhibitors of the cyclin-dependent protein kinases during the yeast cell cycle. On the other side, mass spectrometry is applied to the direct investigation of non-covalent interactions in intact protein structures for conformational studies and binding analysis. Finally, model proteins and peptides are employed for the investigation of the mechanism of electrospray ionization and for the study of non-covalent interactions in the gas phase.
Partecipants: Grandori Rita
STRUCTURE, PHYSIOLOGICAL ROLE AND PATHOGENIC POTENTIAL OF AMYLOIDOGENIC PROTEINS
A wide set of neurodegenerative diseases are related to misfolding and aggregation of some proteins, which eventually results in deposition of amyloids, either intra- or extracellularly, depending on the pathology. The intracellular protein ataxin-3 is responsible for spinocerebellar ataxia type 3, a neurodegenerative disease triggered when the length of a stretch of consecutive glutamines close to the C-terminus of the molecule exceeds a critical threshold, typically 40 residues, with resulting amyloid deposition. To better understand the physiological role(s) of ataxin-3, we are searching for its molecular interactors by taking advantage of proteomic technologies. Also, we are performing investigations on the molecular mechanisms of amyloid fibril formation. To this end, we are characterizing different variants of ataxin-3, carrying either normal or expanded polyglutamine stretches, by circular dichroism, FT-IR and atomic force microscopy.
Partecipants: Fusi Paola Alessandra , Regonesi Maria Elena , Tortora Paolo
room 4051, IV floor, building U3
tel: +39 02 6448 3430