Barbara Spolaore

Department of Pharmaceutical and Pharmacological Sciences, University of Padova
via F. Marzolo 5, 35131 Padova - Italy
Current Position:
Associate Professor of Biochemistry (SSD BIO/10)

Contacts
Phone: +39-049-827-5702
e-mail: barbara.spolaore@unipd.it

  Short CV

Born in Mirano (VE), Italy on the 7th May 1974

1999: Degree summa cum laude in Chemistry and Pharmaceutical Technology, University of Padua

1997: Erasmus student at the Dept. of Organic Chemistry of the University of Wales in Bangor (Wales, UK) in the laboratory of Prof. P.J. Murphy.

1999-2002: PhD in Pharmaceutical Sciences, Dept. of Pharmaceutical and Pharmacological Sciences, University of Padua (Supervisor: Prof. A. Fontana).

2002Visiting scientist at the Dept. of Chemistry, University of Cambridge (UK), laboratory of Mass Spectrometry directed by Prof. C.V. Robinson.

2003-2008: Post-doctoral Fellow at the “Centro di Ricerca Interdipartimentale per le Biotecnologie Innovative” (CRIBI), University of Padua (Supervisor: Prof. A. Fontana).

2003: Visiting scientist at the Dept. of Chemistry, University of Cambridge (UK), laboratory of Mass Spectrometry directed by Prof. C.V. Robinson.

2005: European doctorate in biotechnology (EDBT) awarded by the European Association for Higher Education in Biotechnology (HEduBT).

2008-2022: Assistant Professor of Biochemistry (SSD BIO/10)

2014: Italian National Academic Qualification as Associate Professor of Biochemistry (SSD BIO/10)- call 2012

2022- : Associate Professor of Biochemistry (SSD BIO/10)

Teaching

  Office hours

Upon appointment

  Availability for Thesis Projects

Sono disponibili posti di internato di Tesi di Laurea Magistrale per studenti di CTF, Farmacia e Pharmaceutical Biotechnologies. Il progetto di Tesi verterà sullo studio della conformazione e dinamica di proteine allo scopo di chiarire variazioni conformazionali che causano determinati stati patologici. Le tecniche utilizzate nel progetto riguarderanno la caratterizzazione chimica delle proteine mediante RP-HPLC e spettrometria di massa e la caratterizzazione conformazionale di proteine mediante tecnica di scambio idrogeno-deuterio e spettrometria di massa e mediante tecniche spettroscopiche (dicroismo circolare e fluorescenza).

Research

  Scientific Activities

During the PhD and the post-doctoral period, I worked on the conformational characterization of proteins by means of spectroscopic techniques and by combining limited proteolysis and mass spectrometry. Our group studied the conformational changes determined on model proteins and on proteins of pharmaceutical interest by particular solvent conditions and by the interaction with ligands or fatty acids. 

My scientific interests are still in the field of the chemical and conformational characterization of proteins, mainly by exploiting mass spectrometry techniques. Recently,  I focused on microbial transglutaminase (TGase), an enzyme that allows the site-specific derivatization of proteins at the level of Lys or Gln residues often without affecting protein structure and function. We demonstrated that the conformational features of the protein substrate determine the localization of the sites of TGase derivatization. We are now using this enzyme to modify proteins with fluorophores, chelating agents or other compounds to enable a range of biotechnological applications. 

Currently, we are implenting the hydrogen-deuterium exchange mass spectrometry technique (HDX-MS)  that allows mapping of protein conformation in solution under physiological conditions using small amounts of material. We apply this methodology on different protein systems of physio-pathological relevance, that are under study in our laboratory or in on-going collaborations.

  Technical expertise

Electrospray mass spectrometry (MS) and LC-MS/MS applied to the analysis of the primary structure of proteins.

Analysis of protein structure and dynamics by MS using hydrogen-deuterium exchange mass spectrometry (HDX-MS) and limited proteolysis (Lip-MS). 

Spectroscopic techniques: UV-visible spectroscopy, circular dichroism, fluorescence.

Techniques useful for protein purification and characterization: chromatographic techniques,  gel electrophoresis, chemical and enzymatic hydrolysis of proteins in solution or in gel.

Derivatisation of proteins by chemical reactions and site-specific conjugation using the enzyme transglutaminase. 

  Publications

 

Spolaore, B., Raboni, S., Ramos Molina, A., Satwekar, A., Damiano, N., and Fontana, A. (2012) Local Unfolding Is Required for the Site-Specific Protein Modification by Transglutaminase. BIOCHEMISTRY 51, 8679-8689.

Spolaore, B., Damiano, N., Raboni, S., and Fontana, A. (2014) Site-specific derivatization of avidin using microbial transglutaminase. BIOCONJUG. CHEM. 25(3): 470-480.

Spolaore, B., Raboni, S., Satwekar, A.A., Grigoletto, A., Mero, A., Montagner, I.M., Rosato, A., Pasut, G., and Fontana, A. (2016) Site-Specific Transglutaminase-Mediated Conjugation of Interferon α-2b at Glutamine or Lysine Residues. BIOCONJUG. CHEM. 27(11): 2695-2706.

Antoniel, M., Jones, K., Antonucci, S., Spolaore, B., Fogolari, F., Petronilli, V., Giorgio, V., Carraro, M., Di Lisa, F., Forte, M., Szabó, I., Lippe, G., and Bernardi, P. (2018) The unique histidine in OSCP subunit of F-ATP synthase mediates inhibition of the permeability transition pore by acidic pH. EMBO REP. 19(2): 257-268.

Salvarese, N., Spolaore, B., Marangoni, S., Pasin, A., Galenda, A., Tamburini, S., Cicoria, G., Refosco, F., and Bolzati, C. (2018) Transglutaminase-mediated conjugation and nitride-technetium-99m labelling of a bis(thiosemicarbazone) bifunctional chelator. J. INORG. BIOCHEM. 183: 18-31. 

Spolaore, B., Forzato, G., and Fontana, A. (2018) Site-specific derivatization of human interferon β-1a at lysine residues using microbial transglutaminase. AMINO ACIDS 50(7): 923-932.

Massimino, M.L., Simonato, M., Spolaore, B., Franchin, C., Arrigoni, G., Marin, O., Monturiol-Gross, L., Fernández, J., Lomonte, B., and Tonello, F. (2018) Cell surface nucleolin interacts with and internalizes Bothrops asper Lys49 phospholipase A2 and mediates its toxic activity. SCI REP. 8(1): 10619. 

Spolaore, B., Fernández, J., Lomonte, B., Massimino, M.L., and Tonello, F. (2019) Enzymatic labelling of snake venom phospholipase A(2) toxins. TOXICON 170: 99-107.

Peterle, D., Pontarollo, G., Spada, S., Brun, P., Palazzi, L., Sokolov, A.V., Spolaore, B., Polverino de Laureto, P., Vasilyev, V.B., Castagliuolo, I., De Filippis, V. (2020) A serine protease secreted from Bacillus subtilis cleaves human plasma transthyretin to generate an amyloidogenic fragment. COMMUN. BIOL. 3(1):764.

Bolzati, C., Spolaore, B. (2021) Enzymatic Methods for the Site-Specific Radiolabeling of Targeting Proteins. MOLECULES 26(12):3492.

Ceschi, S., Berselli, M., Cozzaglio, M., Giantin, M., Toppo, S., Spolaore, B., Sissi, C. (2022) Vimentin binds to G-quadruplex repeats found at telomeres and gene promoters. NUCLEIC ACIDS RES. 50(3):1370-1381.

Bolzati, C., Salvarese, N., Spolaore, B., Vittadini, A., Forrer, D., Brunello, S., Ghiani, S., Maiocchi, A. (2022) Water-Soluble [Tc(N)(PNP)] Moiety for Room-Temperature 99mTc Labeling of Sensitive Target Vectors. MOL. PHARM. 19(3):876-894.

Fongaro, B., Cappelleto, E., Sosic, A., Spolaore, B., Polverino de Laureto, P. (2022) DOPET and DOPAC affect the aggregation process of E46K variant of α-synuclein at different extent: insights into the interplay between protein dynamics and catechol effect. PROTEIN SCIENCE 31(7):e4356. 

  Patents

Title: "Method for Labeling of Sensitive and Thermosensitive Targeting Biomolecules with Technetium based Compounds". Inventor(s): Bolzati Cristina; Salvarese Nicola; Refosco Fiorenzo; Ghiani Simona; Maiocchi Alessandro; Spolaore Barbara. Applicant(s):   BRACCO IMAGING SPA [IT]; CONSIGLIO NAZIONALE RICERCHE [IT]. International Publication Number: WO/2018/109164

  Research projects and Funds

2022-2026: AIRC Investigator Grant 2021 (Participant),  “Vimentin-G-quadruplex repeats complexes as innovative therapeutic target to control cancer progression”, PI Prof.ssa Claudia Sissi 

2019-2024: Bando Ricerca Scientifica di Eccellenza – Fondazione Cariparo (Participant), “The Role of Bacterial Proteases in Thrombosis and Amyloidosis -BPiTA”, PI Prof. Vincenzo De Filippis 

2019-2021: PRID-2019 of University of Padua (Participant),  “Interaction between intermediate filament proteins and G-rich oncogene promoters: a novel perspective to control gene expression”, PI Prof. Claudia Sissi 

2018-2020: PRID-2018 of University of Padua (Principal investigator), "Application of Hydrogen Exchange Mass Spectrometry in Molecular Medicine - Probing Protein Structure and Dynamics in Protein Systems of Biopharmaceutical Interest"

2017-2019 PRID-2017 of University of Padua (Participant), “Insight into the inhibition mechanism of α-synuclein aggregation by oleuropein aglycone: a structural point of view”, PI Prof. Patrizia Polverino de Laureto 

2015-2017 PRAT-2015 of University of Padua (Participant), “Mass spectrometry analysis of chemical modifications of proteins involved in thrombotic and rheumatic diseases”, PI Prof. Vincenzo De Filippis