Institution: IBB
Departement: DSB
Typology: Academic collaboration, Facility, Skill, Software
Location: Via Pietro Cstellino 111, 80131 Napoli
Contacts:
- Rita Berisio (rita.berisio@cnr.it)
- Alessia Ruggiero (alessia.ruggiero@cnr.it)
Description:
SKILLS AND COLLABORATIONS The team (see above) uses a wide range of methodologies to unravel complex biology problems, including molecular biology, x-ray crystallography, biophysics, and computational work (molecular modelling and dynamics, docking). The Team actively cooperates with microbiologists, immunologists and cell biologists (e.g. Imperial college London, UCD Dublin, Munich University, Harvard University, Karlsruhe Institute of Technology) and is involved in PNRR projects PE13 INF-ACT, CN1 and CN3. INSTRUMENTATION The molecular biology Labs of this team (Labs 8 and 9) are located in Via Pietro Castellino 111, Naples, and are equipped with (non-exhaustive list) T 100 Thermal Cycler (BIO-RAD), Fume cupboard (Bicasa), Protein Electrophoresis Equipment (BIO-RAD), Shakers Innova S44i (EPPENDORF), Heater at 37°C (G-Therm 075 Carlo Erba), Centrifuge Avanti J-26S XP (Beckman Coulter), Bench Centrifuges (Thermo Scientific), ÄKTA pure™ protein purification system (Cytiva), Autoclave, Sonicator 3000 (Misonix), Microfluidizer (LM20 Microfluidizer® MicrofluidicsTM) The Team has free access to a complex platform for integrated structural biology, acquired by CNR-IBB through several Infrastructural projects, including POR CIRO (2018-2022), PON IMPARA (2019-2023), and the PNRR Infrastructural project SEE-LIFE (StrEngthEning the ItaLIan InFrastructure of Euro-bioimaging, 2023-2025), built to enforce the European network EUROBIOIMAGING. These projects have empowered CNR-IBB by making it a centre of high scientific competence which will be made available to the scientific community. In detail: Protein characterisation Spectrometer UV/Vis (Nanodrop ThermoFisher) Circular Dichroism Spectrophotometer J-1500 (Jasco) Dynamic Light Scattering (Zetasizer Pro Malvern Panalytical) Size-exclusion chromatography (ÄKTA pure™ Cytiva) connected to MiniDAWN Treos spectrometer (Wyatt Instrument Technology) X-ray Crystallography Liquid handling system per 96-well plate (Apricot S1 SPT lab) Automatic crystallising platform (Mosquito® Xtal3 SPT lab) X-ray diffractometer (XtaLAB Synergy-S Rigaku) equipped with HyPix detector. This third-generation source is designed to maximize X-ray photons at the sample by using a combination of new optics, new longer-life tubes and an improved alignment system. The machine is also equipped with microscopes (AZ100 Nikon; Stemi 2000-C Zeiss) Software for solving and visualising structures: HKL3000, XDS, CCP4, Phenix, Pymol Protein-protein and protein-ligand interactions Isothermal titration calorimetry (Microcal PEAQ-ITC Malvern Panalytical) Biolayer interferometry (Octet® R8 BLI System Sartorius) MicroScale Thermophoresis (Monolith and Monolith NT.115 device NanoTemper) Multimode plate reader (EnVison 2105 Revvity) Mass spectrometers for protein characterisation
Team:
- Rita Berisio, Research Director, rita.berisio@cnr.it Long experience (>160 papers) on structural biology and biophysical approaches to unravel human pathologies. Coordinator of the structural biology section of the Italian Crystallography Association. Mentor of EMBL Alumni association (2023-). Research coordinator of EU project BactiVax (2019-24). Trainer di EU COST ENOVA (2019-2022). Coordinator of “DELETE_COVID”. FISR2020IP_00014 (2021-22); of PRIN2022 TENET (2024-26), RECOVER_COVID (2020-21), PRIN 2017SFBFER (2019-24). IBB Responsible of the PNRR project INF-ACT (2023-25). 1 Patent with Max Planck Institute, 2 with the University of Munich.
- Alessia Ruggiero, Researcher, alessia.ruggiero@cnr.it Protein engineering, biochemistry, cloning expression, purification and crystallisation. X-ray crystallography, biophysics (CD, fluorescence, ITC calorimety, thermoforesis, interferometry (BLI) (~100 papers). Coordinator of PRIN 2022 PNRR FIGHT_TB (Prot. P2022JE8FN, 2023-2025). CNR PI of TURBO-Turbidity based Covid-19 test (FISR2020IP_02416, 2021-2022); ii) Block Allocation Group-BAG, presso il sincrotrone Diamond UK.
- Maria Romano, Researcher, maria.romano@cnr.it Protein engineering, expression and purification of membrane proteins, biophysics (~30 papers). Member of Marie Curie Alumni Association (MCAA) (2024-2026); previously member of the council ’Italy Chapter of MCAA (2021-2024). PI of EU project MSCA H2020 - Individual Fellowships (2021-2023). Coordinator of “Cristallograficamente” funded by AIC. Coordinator of PRIN 2022 PNRR ENIGMA-TB (2023-2025). Participant of the CN3 PNRR National Centre.
- Flavia Squeglia, Researcher, flavia.squeglia@cnr.it Molecular biology and biochemistry, crystallisation, biophysical characterisation of proteins and protein-protein interactions by CD, fluorescence MST, ITC, BLI (>50 papers). EMBO short term fellow 2014. Coordinator of PRIN 2020 CANNOT-ESKAPE” (Prot. 2020XNFH9R, 2021-2024). CNR responsible of PRIN 2022 DEFENCE", (Prot. 2022TLZRXT, 2023-2025).
- Valeria Napolitano, TD Researcher PNRR INF-ACT, valeria.napolitano@cnr.it Structural biology and structure-based drug design, MST, ITC, fluorescence polarization, AlphaScreen, NMR) (>20 papers). ESR in AEGIS Marie Skłodowska-Curie ITN (2016-2019) at Jagiellonian University (Krakow, PL) and post-doc at Helmholtz Munich (DE, 2019-2023). Responsible for beamtime allocation at Swiss light source. 1 Patent with Helmholtz Munich
- Luigi Vitagliano, Research Director, luigi.vitagliano@cnr.it Long experience (> 210 papers) on the characterisation of biomacromolecules using both experimental and computational approaches. Editorial member of six International journals, including Scientific reports and Plos One. Currently, he is the national coordinator of a PRIN2022 PNRR (TRA2TRA) and unit coordinator in a PRIN2020 (MOYFLY). Teacher of the Ph.D. doctorate “Biomolecular Sciences” of the Università della Campania L. Vanvitelli.
- Luciana Esposito, Senior researcher, luciana.esposito@cnr.it Long experience (~70 papers) in structure/function/stability of proteins through x-ray crystallography, biophysics and computational studies (MD, docking, modelling, data mining). More recent experience in theoretical studies of nucleic acid sequences adopting a variety of different structures.
- PhD students and research fellows: Giovanni Barra, PhD student; Ornella Ghilardi, PhD student; Mario Privitera, PhD student; Viviana Perrotta, research fellow; Francesco Cinquegrana, research fellow
Figure 1. Biolayer interferometry (Octet® R8 BLI System Sartorius) Figure 2. Liquid handling system per 96-well plate (Apricot S1 SPT lab) Figure 3. Automatic crystallising platform (Mosquito® Xtal3 SPT lab) Figure 4. X-ray diffractometer (XtaLAB Synergy-S Rigaku) equipped with HyPix detector and microscope Stemi 2000-C Zeiss Figure 5. Screening platform to test crystals directly from the xtallisation plate Figure 6. A scheme of the Integrated Structural Biology platform at BMS@IBB