AME – Artificial Metallo Enzymes

Prof. Vincenzo Pavone

Room 2Mc-33; phone +39-081-674399 Email vincenzo.pavone@unina.it

https://www.docenti.unina.it/vincenzo.pavone

https://orcid.org/0000-0001-6432-0802

Prof. Angelina Lombardi

Room 2Mc-35; phone +39-081-674418 Email angelina.lombardi@unina.it

https://www.docenti.unina.it/angelina.lombardi

https://orcid.org/0000-0002-2013-3009

Prof. Flavia Nastri

Room 2Mc-38; phone +39-081-674419 Email flavia.nastri@unina.it

https://www.docenti.unina.it/flavia.nastri

http://orcid.org/0000-0002-3390-9822

 

Dr. Ornella Maglio

Room 2Mc-26; phone +39-081-674400 Email ornella.maglio@unina.it

https://orcid.org/0000-0002-7829-1907

Dr. Marco Chino

Room 2Mc-23; phone +39-081-674421 Email marco.chino@unina.it;

https://www.docenti.unina.it/marco.chino

https://orcid.org/0000-0002-0436-3293

Dr. Daniele D'Alonzo

Room 2Mc-28; phone +39-081-674429 Email daniele.d'alonzo@unina.it;

https://orcid.org/0000-0002-2857-8600

 

Other Group members:

PhD students: Dr. Linda Leone, Dr. Gerardo Zambrano, Dr. Diaa Aref, Dr. Fabio Pirro

Research: Our research focuses on the design of artificial metalloproteins and small molecules with the aim to: (i) master the principles of protein folding and function; (ii) understand the structure-function relationship in metal catalysis; (iii) develop new molecules/materials as drugs or biosensing. Main themes are:

Development of Artificial Metalloenzymes

Design of peptides and peptidomimetics as drugs

Development of bio-inspired devices by immobilization of peptides and proteins on different surfaces

Development of Artificial Metalloenzymes

Enzymes catalyze a wide variety of chemical and biological reactions with high efficiency and selectivity. Metal cofactors are involved in several key steps of life machinery. Metalloproteins regulate redox processes and efficiently catalyze difficult reactions under mild conditions. How the fine-tuning of the scaffolds imparts the wide spectrum of reactivity is of crucial interest both in the fields of structural biology and bioinorganic chemistry. Inspired by natural metalloenzymes, our lab uses de novo design and miniaturization to investigate the minimal requirements for function. Both structural and spectroscopic methods help us to test the success of our designs. Advanced chromatographic and stopped-flow methods support our mechanistic and catalytic studies. Final goal is to engineer artificial metalloenzymes having impact in chemical and biotechnological applications.

Main collaboration:

Prof. W.F. DeGrado (UCSF, USA), Prof. K. Bren (University of Rochester, NY, USA)

Prof. R. O. Louro, (ITQB, New University of Lisbon, Spain)

Prof. V. Balland (Université Paris Diderot, Paris, France)

 

Selected publications:

  • Chino, M., Leone, L., Zambrano, G., Pirro, F., D'Alonzo, D., Firpo, V., Aref, D., Lista, L., Maglio, O., Nastri, F., Lombardi, A. ‘Oxidation Catalysis by Iron and Manganese Porphyrins within Enzyme‐like Cages' Biopolymers (2018), e23107.
  • Zhang, S.-Q., Chino, M., Liu, L., Tang, Y., Hu, X., DeGrado, W. F., Lombardi A., ‘De Novo Design of Tetranuclear Transition Metal Clusters Stabilized by Hydrogen-Bonded Networks in Helical Bundles' J. Am. Chem. Soc. (2018), 140, 1294–1304.
  • Chino, M., Leone, L., Maglio, O., D'Alonzo, D., Pirro, F., Pavone, V., Nastri, F., Lombardi, A., ‘A De Novo Heterodimeric Due Ferri Protein Minimizes the Release of Reactive Intermediates in Dioxygen-Dependent Oxidation' Angew. Chem. Int. Ed. (2017), 56, 15580–15583.
  • Nastri, F., Chino, M., Maglio, O., Bhagi-Damodaran, A., Lu, Y., Lombardi A., ‘Design and Engineering of Artificial Oxygen-Activating Metalloenzymes' Chem. Soc. Rev. (2016), 45, 5020–5054.
  • Vitale, R., Lista L., Cerrone C., Caserta G., Chino, M., Maglio, O., Nastri, F., Pavone, V., Lombardi A., ‘Artificial Heme-Enzyme with Enhanced Catalytic Activity: Evolution, Functional Screening and Structural Characterization' Org. Biomol. Chem. (2015), 13, 4858–4868.

Design of peptides and peptidomimetics as drugs

The development of molecular tools able to freeze linear and cyclic peptides in a well-defined three-dimensional structure, represents a basic research topic for the design of new molecules capable of reproducing the chemical and structural properties of natural systems. They contribute to a better understanding of the structural requirements for function in biological macromolecules, and also allow the development of new, pharmacologically interesting products. Our lab adopts a series of "molecular tools", developed in the last 20 years of experience, to design and synthesize biologically active peptides. These compounds are valuable for the study of biological processes and some of them find applications as drugs (potent inhibitors, agonists, etc.).

Main collaboration:

Dr.ssa M.V Carriero (Istituto Nazionale dei Tumori di Napoli)

Prof. C. Schiraldi (Università degli Studi della Campania "Luigi Vanvitelli")

Dr.ssa A. Arciello

Dr. E. Notomista (Università degli Studi di Napoli "Federico II")

Selected publications:

  • Carriero, M.V., Bifulco, K., Minopoli, M., Lista, L., Maglio, O., Mele, L., Di Carluccio, G., De Rosa, M., Pavone, V. ‘UPARANT: A urokinase receptor-derived peptide inhibitor of VEGF-driven angiogenesis with enhanced stability and in vitro and in vivo potency' Mol. Cancer Ther. (2014), 13, 5 1092-1104.
  • Pavone, V., Zhang, S-Q., Merlino, A., Lombardi, A., Wu, Y., DeGrado, W.F. ‘Crystal structure of an amphiphilic foldamer reveals a 48-mer assembly comprising a hollow truncated Octahedron' Nature Comm. (2014), 5, 3581.
  • Boccella, S., Panza, E., Lista, L., Belardo, C., Ianaro, A., De Rosa, M., de Novellis, V. ‘Preclinical evaluation of the urokinase receptor-derived peptide UPARANT as an anti-inflammatory drug' Inflammatory research (2017), 66, 8 701-709.
  • Gaglione, R., Dell'Olmo, E., Bosso, A., Chino, M., Pane, K., Ascione, F., Itri, F., Caserta, S., Amoresano, A., Lombardi, A., Haagsman, H.P., Piccoli, R., Pizzo, E., Veldhuizen, E.J.A., Notomista, E., Arciello, A. ‘ Novel human bioactive peptides identified in Apolipoprotein B: Evaluation of their therapeutic potential' Biochemical Pharmacology (2017), 130, 34-50.
  • Cimini, D., Corte, K.D., Finamore, R., Andreozzi, L., Stellavato, A., Pirozzi, A.V.A., Ferrara, F. Formisano, R., De Rosa, M., Chino, M., Lista, L., Lombardi, A., Pavone, V., Schiraldi, C. ‘Production of human pro-relaxin H2 in the yeast Pichia pastoris' BMC Biotechnology (2017), 17:4.

Development of bio-inspired devices by immobilization of peptides and proteins on different surfaces

Biomaterials and biodevices are currently changing the biotech and pharma industries. Many start-up all over the world are trying to make this transition possible. We adopt our designed biomolecules (Artificial Metalloenzymes, Bioactive Peptides) to build hybrid nanostructures for diagnostics, bioremediation, and energy storage. By several conjugation methods, we functionalize polymeric surfaces, resins and gold nanoparticles with biological recognition elements, oxidative enzymes or photosensitizers.

Main collaboration:

Dr. R. Battaglia, Dr. M. Mauro (Novaetech S.r.l., Napoli)

Prof. C. De Rosa, Prof. F. Auriemma (Università degli Studi di Napoli "Federico II", Napoli)

Dr. G. Manco, Dr. F. Febbraio (Institute of Protein Biochemistry, CNR, Napoli)

Selected publications:

  • Vitale, R., Lista, L., Lau-Truong, S., Tucker, R.T., Brett, M.J., Limoges, B., Pavone, V., Lombardi, A., Balland, V., ‘Spectroelectrochemistry of FeIII- and CoIII- mimochrome VI artificial enzymes immobilized on mesoporous ITO electrodes' Chemical Communications (2014), 50, 1894-1896.
  • Engelen, M., Lombardi, A., Vitale, R., Lista, L., Maglio, O., Pavone, V., Nastri, F. ‘Branched porphyrins as functional scaffolds for multisite bioconjugation' Biotechnology and Applied Biochemistry (2015), 62, 383-392.
  • Maglio, O., Costanzo, S., Cercola, R., Zambrano, G., Mauro, M., Battaglia, R., Ferrini, G., Nastri, F., Pavone, V., Lombardi, A. ‘A Quartz Crystal Microbalance Immunosensor for Stem Cell Selection and Extraction', Sensors (2017), 17, Article number 2747.
  • Auriemma, F., De Rosa, C., Malafronte, A., Di Girolamo, R., Santillo, C., Gerelli, Y., Fragneto, G., Barker,R., Pavone, V., Lombardi, A. ‘Nano-in-Nano Approach for Enzyme Immobilization Based on Block Copolymers' ACS applied materials & interfaces (2017), 9, 29318-29327.
  • Carullo, P., Chino, M., Cetrangolo, G.P., Terreri, S., Lombardi, A., Manco, G., Cimmino, A.
  • Febbraio, F. 'Direct detection of organophosphate compounds in water by a fluorescence-based biosensing device' Sensors and Actuators, B: Chemical (2018), 255, 3257-3266.

News

  • (March 2018) Marco Chino on Ateneapoli: An interview to Marco Chino describes his hidden passion for the tennis court. (http://www.ateneapoli.it/news/lezioni-di-chimica-tra-i-set-il-tennis-visto-dal-prof-marco-chino)
  • (February 2018) New paper published on Biopolymers: Our paper reviewing the last 20 years of porphyrin-based functional models is now available online. (https://onlinelibrary.wiley.com/doi/abs/10.1002/bip.23107)
  • (January 2018) New paper published on JACS: Our paper describing how we designed a small protein binding an abiotic tetrazinc cluster is now available online. (https://pubs.acs.org/doi/10.1021/jacs.7b08261)
  • (December 2017) New paper published on Angewandte Chemie: Our paper describes the design of a "clicked" diiron metalloenzyme model, the first artificial protein to oxidize the substrate without release of any reactive oxygen species. (https://onlinelibrary.wiley.com/doi/abs/10.1002/anie.201707637)
  • (September 2017) Poster session winner at the SCI National Congress: Our poster on the design of a dicopper metalloenzyme mimicking tyrosinase won the 2017 EurJIC/SCI Poster Session Award assigned by the Inorganic Chemistry Division of Italian Chemical Society, during the SCI National Congress in Paestum (September 2017). (https://drive.google.com/open?id=0B97cPOgTj28MNFNDTzJYUVhjeGhYZmh6eFBlTy1FYUV5NHIw)
  • (May 2017) Prima Scuola Nazionale sui Sensori Chimici: The AME group organized the "First National School on Chemical Sensors" at the Department of Chemical Sciences, University of Naples "Federico II". This event, addressed to PhD students, post-doc and young research associates from Universities, public and private R&D institutes, represented the first school organized by the Interdivisional Group of Sensors that encompasses the Analytical Chemistry, the Electrochemistry, and the Inorganic Chemistry divisions of the Italian Chemical Society. (http://www.soc.chim.it/it/gruppi/sensori/Scuola2017)
  • (May 2014) Biophysics of Amyloids and Prions: A. Lombardi and F. Nastri organized in Naples the Biophysics of Amyloids and Prions international meeting. This meeting was one of the event organized on the occasion of the "Year of the Italian Culture in the USA", under an agreement for co-operation in the field of scientific research between Federico II University and the University of California San Francisco. The meeting aimed at covering the emerging field of biophysical approaches to understanding amyloid and prion formation, and deciphering the structural basis for self-propagating protein conformations. (http://2ubythebay.org/). The scientific level of the meeting was of considerable importance given the participation of Professor Stanley Prusiner (UCSF), Nobel Prize for Medicine in 1997 for his discovery of prions.