Del Giudice Alessandra

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1 1 Del Giudice Alessandra address: Mobile: Telephone (Home): Personal Information Date of birth: 29/10/1988 Place of birth: San Severo (Foggia) - Italia Address: Via di Sant'Ippolito, Roma - Italia Citizenship: Italian Current position From March 2015: Visiting PhD student at the Division of Pure and Applied Biochemistry, LTH Lund, Sweden. From October 2013: PhD student in Chemical Sciences, at Department of Chemistry, Sapienza University of Rome. Field: Physical Chemistry, structural and spectroscopic studies on Soft Matter systems. Provisional thesis title: "Structural and spectroscopic studies on proteins in stress conditions". Supervisor: Prof. Nicolae Viorel Pavel. Scientific activities Peer-reviewed publications Conference talks 1. Ibuprofen and Propofol Cobinding Effect on Human Serum Albumin Unfolding in Urea Alessandra Del Giudice, Claudia Leggio, Nicole Balasco, Luciano Galantini, and Nicolae V. Pavel The Journal of Physical Chemistry B (34), Structural insights into the shape and assembly of photosynthetic GAPDH/CP12/PRK complex by small angle X-ray scattering Alessandra Del Giudice, Simona Fermani, Francesca Sparla, Paolo Trost, Nicolae Viorel Pavel 2 nd Joint AIC-SILS Conference, Florence, September Research experience April present PhD thesis project Structural and spectroscopic studies on proteins in stress conditions.

2 2 November present The project is aimed to the structural characterization, aided by spectroscopic techniques, of Human Serum Albumin (HSA) damage processes in stress conditions (including unfolding, aggregation, fragmentation). The possible protective effect of HSA ligands (drugs, antioxidants) will be investigated. Processes of chemical, heat and oxidative induced denaturation will be considered. In particular, proteins undergoing oxidative stress can result in modified forms identified in some pathological conditions. Preceding studies in the biomedical field are mainly based on chemical and spectroscopic essays, neglecting dimensional aspects. We would like to characterize the structural evolution by means of small angle X-ray scattering (SAXS) and light scattering, in order to obtain information on the shape and dimension of the species evolving during the damage process. Complementary spectroscopic techniques (fluorescence, UV-vis, circular dichroism) would help to monitor variations in the secondary and tertiary protein structure. Contemporary synchrotron-radiation-saxs and spectroscopic mesaurements are also planned. The developement of proper data-analysis procedures, in particular for the analysis of SAXS data in presence of protein aggregation, and also the use of separative techniques could be necessary. Collaborative project with University of Bologna Small-angle X-ray scattering study of photosyntetic proteins and their regulatory complexes. Calvin cycle enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) form together with the scaffold protein CP12, a supramolecular ternary complex of 498 kda with stoichiometry [(GAPDH)-(CP12)-(PRK)] 2. CP12 is an ubiquitous regulatory protein of oxygenic phototrophs that contains, with few exceptions, four conserved cysteines able to form two consecutive disulfide bridges. In higher plants as Arabidopsis thaliana, CP12 is predicted to be an intrinsically disordered protein (IDPs). The activities of GAPDH and PRK enzymes are inhibited by complex formation and fully recovered upon dissociation of the complex at the onset of light, providing an effective means for regulation of the Calvin cycle in vivo. It is proposed that GAPDH/CP12/PRK supramolecular complex occurs in chloroplasts in the dark to ensure strong down-regulation of the Calvin cycle. Thus, the determination of the ternary complex structure is crucial for the understanding of the photosynthetic metabolism in light/dark regime. Crystallization trials to produce single crystals of the complex for X- ray diffraction experiments, failed. A structural study in solution by small angle scattering was then approached. May July 2013 Master's degree thesis project Denaturation and stabilization of Human Serum Albumin: combined effect of

3 3 drugs. The denaturation path of Human Albumin at increasing urea concentrations has been previously studied with Small Angle X-Ray Scattering, light scattering and spectroscopic techniques, for the ligand-free protein, the palmitic acid-bound protein, as well as in presence of Ibuprofen, thus outlining a mechanism for the subsequent unfolding of the protein domains and the protective effect of ligands. In this work, the anesthetic drug Propofol was added to the HSA-Ibuprofen system in order to assess the combined effect of the drugs (that have a common binding site on the protein, along with other secondary sites). The complete SAXS data set at increasing denaturant concentrations was analyzed with the Singular Value Decomposition method, thereby allowing the presence of two intermediates along with the Native and the Unfolded state to be identified. The 3D structures at low resolution of the intermediates were restored from the scattering data, and this led to a comparison with the previously determined unfolding mechanism in absence of Propofol; its presence seems to stabilize compact forms of the protein even at high denaturant concentrations. October April 2011 Bachelor's degree thesis project The effect of ph on the stability of Human Serum Albumin: spectroscopic investigations. HSA is known to undergo conformational transitions with varying ph and, along with the Native form stable around neutral ph, the existence of other isomers is reported in the literature. The aim of my work was to investigate the behavior of the protein in a broad range of ph by means of two spectroscopic techniques: static fluorescence and circular dichroism. I also analyzed far-uv dichroic spectra with the package CD-Pro, while fluorescence spectra were subjected to second derivative analysis. Measurements were repeated in order to obtain reproducible trends. It was a preparatory study designed to further characterize the Albumin isomers by means of SAXS and light scattering. Education 2 nd level degree Period: From 1/10/2011 to 18/07/2013 University: Sapienza Università di Roma Name of the degree: Master's Degree in Chemistry Final degree mark: 110/110 cum laude

4 4 Thesis title: Advisor: Subject area of study: Denaturation and stabilization of Human Serum Albumin: combined effect of drugs. Prof. Nicolae Viorel Pavel Physical Chemistry Additional information Specific field of the course: Master's Degree in Chemistry, Inorganic-Physical Chemistry Course ("Indirizzo Inorganico-Chimico Fisico, ICF") Principal subjects: Thermodynamics; spectroscopic and structural techniques; computational chemistry; preparation and physical-chemical and structural characterization of inorganic and organic materials. Average grade: 30/30 Website of the degree course: 1 st level degree Period: From 1/10/2007 to 4/05/2011 University: Name of the degree: Final degree mark: Thesis title: Advisor: Subject area of study: Sapienza Università di Roma Bachelor's Degree in Chemistry 110/110 cum laude The effect of ph on the stability of Human Serum Albumin: spectroscopic investigations Prof. Nicolae Viorel Pavel Physical Chemistry Additional information Contents: Average grade: 29.5/30 Website of the degree course: Fundamentals of mathematics and physics; wide theoretical preparation in the fields of inorganic, analytical, physical, organic, biological chemistry; some practical training in the fundamental laboratory operations. Pre-academic studies Period: From 1/09/2002 to 20/07/2007

5 5 School: Qualification: School-leaving examination mark: Liceo Scientifico Checchia-Rispoli San Severo /FG), Italy (Public school) Italian scientific secondary school diploma 100/100 cum laude Additional training 23 rd February - 26 th March 2014 Higher European Research Course for Users of Large Experimental Systems (HERCULES School 2014, Session B: applications to biomolecular structure and dynamics Grenoble/Saclay, France Presentation of a poster "Denaturation and stabilization of Human Serum Albumin: combined effect of drugs" 23 rd February - 6 th March th IFF Spring School - Functional Soft Matter Forschungszentrum Jülich, Germany ( e.html) Experimental techniques Main (Practical knowledge, very familiar) Small Angle X-Ray Scattering Circular Dichroism Fluorescence UV-visible spectroscopy Dynamic Light Scattering Foreign language skills Europass self-classification Overall Understanding Speaking Writing Listening Reading English Excellent C1 C1 B2/C1 B2/C1 German Limited A1 A1 A1 A1

6 6 Information technology skills Specific skills Programming languages Fortran 77, Fortran 95 Software applications Data eleaboration and plot creation: Gnuplot Sigmaplot Kaleidagraph Igor TopDrawer Small Angle Scattering data analysis: ATSAS Package - EMBL ITP - Glatter GENFIT - Spinozzi Ab-initio molecular energy calculation: Gamess Didactic-level molecular dynamics: Tinker PDB files visualization and elaboration: PyMOL VMD Chimera ViewerLite Certifications ECDL Awards and scholarships February 2014 Winner of a grant for a 150 hour-collaboration as tutor at Department of Chemistry, Sapienza Univeristy of Rome. from March 2014 to August 2014: tutor for the Physical Chemistry III course (Spectroscopy) October 2013 Successful candidate in an open competition for the assignment of PhD fellowships for the Chemical Sciences PhD Course at Department of Chemistry, Sapienza Univeristy of Rome. First place URL: che_xxix_ciclo-graduatoria_finale.pdf

7 7 November 2011 Winner of a grant for a 150 hour-collaboration as physical chemistry didactic laboratories assistant at Department of Chemistry, Sapienza Univeristy of Rome. from November 2011 to February 2012: 75 hours: laboratory of thermodynamics from October 2012 to January 2013: 75 hours: laboratory of spectroscopy