Personale docente

Full professor in Applied Physics at the Faculty of Medicine, University of Parma.
He is a co-author of about 80 papers on international, peer-reviewed journals, books or book chapters (h-index = 20, Thomson Reuters ISI Web of Knowledge) and more than 100 communications to national and international meetings.

EDUCATION:
1992: graduated cum laude in Veterinary Medicine at the University of Parma, Italy; thesis on “Allosteric regulation of the multifunctional enzyme tryptophan synthase”.
1997-1998: Fellow, Italian National Institute for the Physics of Matter (INFM). Research carried out at the Institute of Physical Sciences and Institute of Biochemical Sciences, University of Parma, on “Folding of O-acetylserine sulfhydrylase by tryptophan and coenzyme steady-state and time-resolved fluorescence”.
1998: Ph.D., Molecular Biology and Pathology, University of Modena, Italy; thesis on “Oxygen binding to hemoglobin in the T quaternary state”. Research carried out at the Institute of Biochemical Sciences, University of Parma, Prof. Andrea Mozzarelli, Supervisor.
1999-2000: Post-doctoral fellow, Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD, USA, Dr. William A. Eaton, Supervisor.

PROFESSIONAL EXPERIENCE
2000-2001: Research contract with the Faculty of Pharmacy of the University of Parma. Research activity on “Mechanisms of protein folding-unfolding processes: structural, dynamic and functional characterization of pyridoxal 5’-phosphate-dependent enzymes and heme proteins”.
2001-2006: Permanent position as a “ricercatore” (similar to assistant professor) in Applied Physics, Faculty of Medicine and Surgery, University of Parma, Italy.
2006 - present: Full Professor, Applied Physics, Faculty of Medicine and Surgery, University of Parma, Italy.

FELLOWSIPS AND AWARDS
1997-1998: Fellowship from the Italian National Institute for the Physics of Matter (INFM); part of work on O-acetylserine sulfhydrylase carried out at University of Illinois at Urbana-Champaign, USA, in collaboration with E. Gratton.
1998: National Prize for Ph.D. Candidates by the Italian Society of Differentiation and Cellular Biology (ABCD).
1999-2000: Long-term fellowship by the European Molecular Biology Organization (EMBO) and Supplemental Postdoctoral Visiting Fellow Award of the NIH, Supervisor: W. A. Eaton.
2003: Prize of Italian Society of Physics for best presentation at the 89th National Meeting in the Biophysics and Medical Physics Section.

SOCIETY MEMBERSHIPS
Italian Physics Society.
CNISM (National Inter-university Consortium for the Physics of Matter).
Italian National Institute of Biostructures and Biosystems.
American Society for Biochemistry and Molecular Biology.

OTHER ACTIVITIES
Reviewer for international journals such as Biochimica et Biochimica Acta – Proteins and Proteomics, Biotechnology & Bioengineering, Protein and Peptide Letters, Journal of Functional Biomaterials.
Guest Editor, with Cristiano Viappiani (University of Parma, Italy) and F. Javier Luque (University of Barcelona, Spain), of a Special Issue on "Protein Dynamics, experimental and computational approaches" of the international journal Biochimica and Biophysica Acta - Proteins and Proteomics (2011).
Co-editor, with Andrea Mozzarelli (University of Parma) of the book “Chemistry and Biochemistry of Oxygen Therapeutics: from Transfusion to Artificial Blood”, John Wiley and Sons, Ltd. (2011).
Member, organizing committee of the International Course “From structural biology to drug discovery”, Parma, September 29, 2000.
Member, scientific and organizing committees of the International Courses “From structural genomics to drug discovery”, Parma, September 27-28, 2002 and September 27-28, 2004.
Member, organizing committee of the International Conference “International Visions on Blood Substitutes. Hemoglobin-Based Oxygen Carriers: from Chemistry to Clinic”, Parma, September 10-20, 2006.
Vice-chairman and member of the organizing committee of the international conference "New Challenges in Protein Science", Parma, June 4-6, Giugno 2008.
Member of the organizing committee of the international conference “XII International Symposium on Blood Substitutes”, Parma, August 25-28, 2009.
Member of Biopharmanet-Tec laboratory (Interdepartmental center for the innovation of health products).
Member of SITEIA (Interdepartmental center for food safety technolgy innovation).

RECENT RESEARCH ACTIVITY
1) Encapsulation of biological macromolecules in wet nanoporous silica gels experienced a boost in the last decade. The research carried out by our group contributed to the development and diffusion of this technique, which allows to immobilize biological macromolecules in an inorganic three-dimensional matrix. This strategy permits: i) to isolate molecular species that are metastable or poorly populated in solution, making them accessible to spectroscopy and microscopy studies also with single molecule resolution, as in the case of Green Fluorescent Protein (GFP); ii) to mimic, in vitro, biologically relevant molecular confinement effects on macromolecules conformational equilibria and dynamics; iii) to develop protein-based biosensors and bioreactors. Experiments on green fluorescent protein (GFP) mutants, in solution and in wet nanoporous silica gels, proved that the structural and dynamic properties are maintained upon silica gel encapsulation. Therefore, encapsulation in silica gel was exploited to investigate, by confocal microscopy, the photochemical and photodynamic properties, the kinetic and thermodynamic stability and the folding mechanism of GFP with single molecule resolution. The dynamics of spectral and structural changes of the GFP chromophore as a function of pH was investigated by measuring transient absorption and fluorescence emission following laser-induced proton release from caged compounds. Experiments on site-specific mutants, carried out in collaboration with Prof. Cristiano Viappiani, University of Parma, contributed to shed light on the structural basis of proton transfer pathways and conformational dynamics controlling the interconversion among protonation states of the chromophore. This topic was also investigated by one- and two-photon excited Fluorescence Correlation Spectroscopy, in collaboration with Drs. Giancarlo Baldini, Giberto Chirico and Maddalena Collini, University of Milan-Bicocca. We are currently investigating, by microspectrophotometry, the spectroscopic and dynamic properties of GFP mutants in the crystalline state. This will allow to directly compare functional properties with the structural information that will be obtained by determining the three-dimensional structure in the same physical state, in collaboration with prof. Roberto Battistutta, University of Padova.
2) The understanding of the mechanism of hemoglobin (Hb) allosteric regulation has been implemented through the correlation of structural data in the presence and absence of ligands with functional properties of immobilized Hb. Several Hbs were investigated: HbA, mutant or chemically modified human Hbs, Fe-Ni metal hybrids (Ni-substituted hemes do not bind oxygen). Part of these studies, initiated during a visit at the Dept. of Medicine, State University of New York at Buffalo, Buffalo, NY, USA, were carried out in collaboration with Prof. Robert W. Noble. The functional properties of Hbs immobilized in a single quaternary state by crystallization or encapsulation in silica gels have been investigated with microspectrophotometric techniques. In the gel, by varying the gelification protocol and/or experimental conditions, it is possible to block or kinetically uncouple the tertiary and quaternary relaxations that follow ligand binding, and are inhibited by crystal lattice. This makes accessible to spectroscopic investigation metastable intermediates and protein conformations that in solution are poorly populated, or normally present in heterogeneous mixtures. Equilibrium and laser flash-photolysis studies of oxygen and carbon monoxide binding to gel-encapsulated HbA highlighted the functional heterogeneity of the T quaternary state. These studies led to the development and validation of a model of allosteric regulation of Hb, representing an extension of Monod, Wyman and Changeux model, with better performance in describing heterotropic ligand effects. The model was developed during the post-doc period (1999-2000) at the Laboratory of Chemical Physics, NIDDK, NIH, Bethesda, MD, USA, directed by Dr. William A. Eaton. During a following visit at the Dept. of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY, USA, Raman spectroscopy experiments on Hb encapsulated in silica gel were carried out in collaboration with Prof. Joel M. Friedman. The experiments, together with circular dichroism spectroscopic studies carried out in Parma, yielded structural information on the different tertiary conformations populating the T quaternary state as a function of experimental conditions. The most recent activity yielded further evidence in favor of the proposed allosteric model, through the characterization of the time evolution of the distribution of tertiary and quaternary species following CO binding and photolysis in the gel.
Ongoing studies are aimed at testing the potential application of chemically or genetically modified Hbs for oxygen transport in vivo.
3) Pyridoxal 5’-phosphate (PLP)-dependent proteins belong to a large enzyme superfamily characterized by high catalytic versatility. PLP-dependent enzymes are classified in three functional families based on the stereochemistry of the catalyzed reactions, and five fold-types based on sequence and structure similarity. Sequence homology is often very low even within the same family or fold type, despite common structural motives and reaction mechanism. Therefore, the investigation of protein dynamics and reaction and folding mechanism of PLP-dependent enzymes provides a rare opportunity to investigate the relationships among sequence, structure, function and folding. The folding mechanism of the B isozyme of O-acetylserine sulfhydrylase (OASS) was characterized by absorption, steady-state and time resolved fluorescence, circular dichroism, nuclear magnetic resonance and dynamic light scattering spectroscopy. Other OASS isoforms were also studied, as well as the structural, dynamic and functional effects of interprotein inter actions with other enzymes involved in sulphur metabolism. We are currently testing peptides and peptidomimetic molecules with inhibitory effects on OASS function, in view of their potential development as antibacterial drugs.
Fluorescence correlation spectroscopy and two-photon excitation time-resolved fluorescence experiments were carried out during several visits at the Laboratory of Fluorescence Dynamics, University of Illinois at Urbana-Champaign, Urbana, IL, USA, directed by Prof. Enrico Gratton (now at University of California at Irvine). The experiments allowed to resolve the molecular heterogeneity of OASS at a single molecule level. New information was achieved on the distribution of enzyme tautomers in the ground and excited states.
The catalytic reaction and allosteric regulation of tryptophan synthase have been investigated through the characterization of the functional properties of site-specific mutants and stopped-flow experiments on the pH-dependence of reaction rates and distribution of catalytic intermediates.

FUNDING HISTORY
1) Coordinator of the Unit of Parma for the PRIN Project (2008) “Development of fluorescent proteins for optical nanoscopy oriented to the study of cellular dynamics" (national coordinator: Prof. A. Diaspro, University of Genova). The Unit of Parma is working on “Design, expression, and spectroscopic and structural characterization of fluorescent proteins for optical micro- and nanoscopic applications”.
2) Coordinator of the Unit of Parma for the PRIN Project (2006) “Conformational substates and folding-unfolding pathways in green fluorescent protein: an experimental and theoretical study towards discrete states in proteins” (national coordinator: Prof. G. Baldini, University of Milan-Biococca). The Unit of Parma worked on “Spectroscopic characterization of GFPmut2 and site-specific mutants in solution and in silica gel, in the native state and under denaturing conditions”.
3) Member of a University of Parma research group funded by the Cassa di Risparmio di Parma Foundation (beginning 2008) for research on “Biotechnological approaches to hypo-oxygenation pathologies: hemoglobin-based blood substitutes”.
4) Member of a group funded by the Italian Ministry of University and Research for an internationalization project, carrying out activity in collaboration with the Virginia Commonwealth University; Richmond, VA, and the University of Oklahoma, Norman, OK, USA.
5) European Union 2004-2007 project within the VI Framework Program, on “EuroBlood Substitutes”, coordinated by Dr. Ken Lowe, University of Nottingham, UK, and locally by Prof. Andrea Mozzarelli.
6) CNR Strategic Project “Functional Genomics” on “Structural basis of evolution and protective activity of the host defensive mechanisms of truncated hemoglobins expressed by unicellular pathogens”, coordinated by Prof. Martino Bolognesi and later by Prof. Andrea Mozzarelli.
7) PRIN Project (2003) on “Molecular basis of pyridoxal 5’-phosphate-dependent enzymes catalytic versatility”. National coordinator, Prof. F. Bossa. The Unit of Parma, coordinated by Prof. Andrea Mozzarelli, worked on “Functional genomics, regulation, stability and biotechnological applications of pyridoxal 5’-phosphate-dependent enzymes”.
8) FIRB Project (2001) on “Molecular nanodevices”. National coordinator, Prof. Roberto Cingolani; local coordinator, Prof. Marco Fontana.
9) PAIS Project (2001-2002) of the National Institute for the Physics of Matter “SINGMOL - Structural and dynamic studies of proteins by single molecule spectroscopies”. Coordinator, Prof. Giuseppe Chirico.
10) PRIN Project (2001) on “Structural organization, plasticity and catalytic versatility of pyridoxal 5’-phosphate-dependent enzymes”. National coordinator, Prof. Carla Borri Voltattorni. The Unit of Parma, coordinated by Prof. Andrea Mozzarelli, worked on “Isolation and characterization of catalytic and folding intermediates in pyridoxal 5’-phosphate-dependent enzymes in the crystal, in silica gel and in solution”.
11) Sub-contract between the University of Parma and the University of New York at Buffalo, within Project 2, coordinated by Prof. Robert W. Noble, for the NIH research project (1999-2003) “Hemoglobin: a workbench to study protein allostery”. Coordinator, Dr. Arthur Arnone, University of Iowa, USA.
12) CNR Project (1996) “Biotechnologies”. Coordinator, Prof. Gian Luigi Rossi.
13) CNR Project (1996, 1997 and 1998) “Regulation and intersubunit interactions in pyridoxal 5’-phosphate-dependent enzymes and in hemoglobin”.
14) NATO Grant (1993-2000) “Oxygen binding to single crystals of hemoglobin”, in collaboration with Dr. William A. Eaton, NIH, Bethesda, MD, USA.