U.O. name

Structural and Cellular Biochemistry

Scientific Coordinator

Prof. Gianluca Damonte

Description

Four main research area, detailed below, are investigated:

• Abscisic acid (ABA): a new endogenous hormone in humans. Our group has demonstrated that ABA is an endogenous hormone in humans, that is released from different cell types and that stimulates a functional response in different cell types. Specifically, ABA stimulates, on one hand, insulin release from beta-pancreatic cells and, on the other, glucose influx in myocytes and adipocytes, thus contributing to the regulation of glucose homeostasis. The ABA-triggered signaling cascade has been described, following its binding to its human receptor, LANCL2, identified by us. We aim at studying ABA role in the physiology of glucose homeostasis regulation and in type 2 diabetes pathogenesis.
• Biosynthesis and metabolism of NAD+ as new therapeutic targets in cancer.. Appropriate levels of nicotinamide adenine dinucleotide (NAD+) are important in the control of various processes. Enzymes involved in NAD+ synthesis and degradation are emerging as possible targets of new therapeutic agents, modulating enzyme activities. In our studies, we will aim at identifying new anti-cancer strategies based on the inhibition of NAD+ synthesis and on the modulation of its metabolism.
• Analysis and characterisation of GDP-L-fucose metabolism and of its uptake and utilization inside the Golgi apparatus. This activity concerns, on one hand, the analysis and characterization of GDP-L-fucose metabolism and uptake into the Golgi system and on the other the identification of the correlation between GDP-L-fucose availability and production of fucosylated glycans. Moreover, the role of fucosylated glycans and physiopathological conditions during development and cell differentiation is investigated. Specific skills of the laboratory include methodologies of molecular and cell biology, production and characterization of recombinant proteins, mutagenesis, structural analysis of glycoconiugates, cell transfection and RNA silencing.
• Structural characterization and qualitative-quantitative analysis of biomolecules and metabolites. This research area regards the structural characterization and qualitative-quantitative determination of proteins, oligonucleotides, carbohydrates, lipids, steroid-like molecules with potential pharmacological activities, e.g. antineoplastic and antiviral drugs and pro-drugs. The characterization and quality control of encapsulated molecules in engineered erythrocytes employed as suitable bioreactors for time-controlled and cell-targeted delivery of cytotoxic drugs represents another characteristic activity. Moreover the synthesis, purification and structural characterization and quality control of creatine derivatives for the study of its transporter "in vivo" and for the therapy of its deficit, as well as the synthesis, characterization and biological application of aminoarylthiazoles for the correction of chloride transport in cystic fibrosis.

List of publications

1. Bruzzone S, Moreschi I, Usai C, Guida L, Damonte G, Salis A, Scarfì S, Millo E, De Flora A, Zocchi E. Abscisic acid is an endogenous cytokine in human granulocytes with cyclic ADP-ribose as second messenger. Proc Natl Acad Sci USA. 2007 Apr 3;104(14):5759-64. Epub 2007 Mar 26. PMID:17389374
2. Bruzzone S, Bodrato N, Usai C, Guida L, Moreschi I, Nano R, Antonioli B, Fruscione F, Magnone M, Scarfì S, De Flora A, Zocchi E. Abscisic acid is an endogenous stimulator of insulin release from human pancreatic islets with cyclic ADP ribose as second messenger. J Biol Chem. 2008 Nov 21;283(47):32188-97. doi: 10.1074/jbc.M802603200. Epub 2008 Sep 10. PMID:18784081
3. Armirotti A, Damonte G, Pozzolini M, Mussino F, Cerrano C, Salis A, Benatti U,Giovine M. Primary structure and post-translational modifications of silicatein beta from the marine sponge Petrosia ficiformis (Poiret, 1789). J Proteome Res. 2009;8(8):3995-4004.PMID: 19522542
4. Sturla L, Fresia C, Guida L, Bruzzone S, Scarfì S, Usai C, Fruscione F, Magnone M, Millo E, Basile G, Grozio A, Jacchetti E, Allegretti M, De Flora A, Zocchi E. LANCL2 is necessary for abscisic acid binding and signaling in human granulocytes and in rat insulinoma cells. J Biol Chem. 2009 Oct 9;284(41):28045-57. doi: 10.1074/jbc.M109.035329. Epub 2009 Aug 10. PMID: 19667068
5. Scapolla C, Cangemi G, Barco S, Barbagallo L, Bugnone D, Maffia A, Melioli G,Profumo A, Benatti U, Damonte G. Identification and structural characterization by LC-ESI-IONTRAP and LC-ESI-TOF of some metabolic conjugation products of homovanillic acid in urine of neuroblastoma patients. J Mass Spectrom. 2012, Jul;47(7):816-24.PMID: 22791248.
6. Bruzzone S, Ameri P, Briatore L, Mannino E, Basile G, Andraghetti G, Grozio A, Magnone M, Guida L, Scarfì S, Salis A, Damonte G, Sturla L, Nencioni A, Fenoglio D, Fiory F, Miele C, Beguinot F, Ruvolo V, Bormioli M, Colombo G, Maggi D, Murialdo G, Cordera R, De Flora A, Zocchi E. The plant hormone abscisic acid increases in human plasma after hyperglycemia and stimulates glucose consumption by adipocytes and myoblasts. FASEB J. 2012 Mar;26(3):1251-60. doi: 10.1096/fj.11-190140. Epub 2011 Nov 10. PMID: 22075645
7. Piacente F, Marin M, Molinaro A, De Castro C, Seltzer V, Salis A, Damonte G, Bernardi C, Claverie JM, Abergel C, Tonetti M. Giant DNA virus mimivirus encodes pathway for biosynthesis of unusual sugar 4-amino-4,6-dideoxy-D-glucose (Viosamine). J Biol Chem. 2012, 27;287(5):3009-18. PMID: 22157758
8. De Castro C, Molinaro A, Piacente F, Gurnon JR, Sturiale L, Palmigiano A, Lanzetta R, Parrilli M, Garozzo D, Tonetti MG, Van Etten JL. Structure of N-linked oligosaccharides attached to chlorovirus PBCV-1 major capsid protein reveals unusual class of complex N-glycans. Proc Natl Acad Sci U S A. 2013, 20;110(34):13956- PMID: 23918378
9. Bernardi C, Soffientini U, Piacente F, Tonetti MG. Effects of microRNAs on fucosyltransferase 8 (FUT8) expression in hepatocarcinoma cells. PLoS One. 2013 9;8(10):e76540. PMID: 24130780.
10. Grozio A, Sociali G, Sturla L, Caffa I, Soncini D, Salis A, Raffaelli N, De Flora A, Nencioni A, Bruzzone S. CD73 protein as a source of extracellular precursors for sustained NAD+ biosynthesis in FK866-treated tumor cells. J Biol Chem. 2013 Sep 6;288(36):25938-49. doi: 10.1074/jbc.M113.470435. Epub 2013 Jul 23. PMID: 23880765
11. Parenti MD, Grozio A, Bauer I, Galeno L, Damonte P, Millo E, Sociali G, Franceschi C, Ballestrero A, Bruzzone S, Del Rio A, Nencioni A. Discovery of novel and selective SIRT6 inhibitors. J Med Chem. 2014 Jun 12;57(11):4796-804. PMID: 24785705
12. Piacente F, Bernardi C, Marin M, Blanc G, Abergel C, Tonetti MG. Characterization of a UDP-N-acetylglucosamine biosynthetic pathway encoded by the giant DNA virus Mimivirus. Glycobiology. 2014;24(1):51-61. PMID: 24107487
13. Vaisitti T, Audrito V, Serra S, Buonincontri R, Sociali G, Mannino E, Pagnani A, Zucchetto A, Tissino E, Vitale C, Coscia M, Usai C, Pepper C, Gattei V, Bruzzone S, Deaglio S. The enzymatic activities of CD38 enhance CLL growth and trafficking: implications for therapeutic targeting. Leukemia. 2015 Feb;29(2):356-68. doi: 10.1038/leu.2014.207. Epub 2014 Jul 3. PMID: 24990614
14. Piacente F, De Castro C, Jeudy S, Molinaro A, Salis A, Damonte G, Bernardi C, Abergel C, Tonetti MG. Giant virus Megavirus chilensis encodes the biosynthetic pathway for uncommon acetamido sugars. J Biol Chem. 2014, 29;289(35):24428-39. PMID: 25035429;
15. De Castro C, Speciale I, Duncan G, Dunigan DD, Agarkova I, Lanzetta R, Sturiale L, Palmigiano A, Garozzo D, Molinaro A, Tonetti M, Van Etten JL. N-linked Glycans of Chloroviruses Sharing a Core Architecture without Precedent. Angew Chem Int Ed Engl. 2015, [Epub ahead of print] PMID: 26582281.
16. Magnone M, Ameri P, Salis A, Andraghetti G, Emionite L, Murialdo G, De Flora A, Zocchi E. Microgram amounts of abscisic acid in fruit extracts improve glucose tolerance and reduce insulinemia in rats and in humans. FASEB J. 2015 Aug 4. pii: fj.15-277731. [Epub ahead of print]. PMID: 26243865
17. Bruzzone S, Magnone M, Mannino E, Sociali G, Sturla L, Fresia C, Booz V, Emionite L, De Flora A, Zocchi E. Abscisic Acid Stimulates Glucagon-Like Peptide-1 Secretion from L-Cells and Its Oral Administration Increases Plasma Glucagon-Like Peptide-1 Levels in Rats. PLoS One. 2015 Oct 21;10(10):e0140588. doi: 10.1371/journal.pone.0140588. eCollection 2015. PMID: 26488296
18. Bellotti M, Salis A, Grozio A, Damonte G, Vigliarolo T, Galatini A, Zocchi E, Benatti U, Millo E. Synthesis, structural characterization and effect on human granulocyte intracellular cAMP levels of abscisic acid analogs. Bioorg Med Chem. 2015, 1;23(1):22-32. PMID: 25496807.
19. Sociali G, Raffaghello L, Magnone M, Zamporlini F, Emionite L, Sturla L, Bianchi G, Vigliarolo T, Nahimana A, Nencioni A, Raffaelli N, Bruzzone S. Antitumor effect of combined NAMPT and CD73 inhibition in an ovarian cancer model. Oncotarget. Accepted: November 16, 2015.
20. Pozzolini M, Scarfì S, Mussino F, Salis A, Damonte G, Benatti U, Giovine M. Pichia pastoris production of a prolyl 4-hydroxylase derived from Chondrosia reniformis sponge: A new biotechnological tool for the recombinant production of marine collagen. J Biotechnol. 2015, Aug 20;208:28-36.PMID: 26022422

Main research lines

• Study of abscisic acid functional effects and related biomolecular mechanism in different human and animal cellsi. Specifically, we will investigate ABA role in the physiology of glucose homesostasis regulation and in the pathogenesis of type 2 diabetes, by evaluating on in vitro and in vivo models, the possible cross-talk between ABA and the incretin GLP-1 and the effect of oral ABA administration in glycemia regulation, in diabetes animal models.
• Structural and functional characterization of the abscisic acid receptor, LANCL-2, that could represent a new pharmacological target in different pathologies; study of the metabolic and behavioural phenotype of potrebbe rappresentare un inedito bersaglio farmacologico in diverse patologie umane; we will study the metabolic and behavioural phenotype of the LANCL2 knock-out mice, recently generated in our lab.
• Identification of antagonist of Sirtuin 6, an enzyme with a NAD+-dependent deacetylase activity and their potential use in the treatment of cancer.
• Decrease of intracellular NAD+ synthesis, through the pharmacological inhibition of the enzymes involved in its synthesis, and evaluation of their anti-tumoral effects in in vitro and in vivo models.
• Analysis and characterization of GDP-L-fucose metabolism and uptake into the Golgi system
• Role of fucosylated glycans during development and cell differentiation.
• Analisi proteomica di molecole enzimatiche e di collagene di organismi marini mediante tecniche di spettrometria di massa (MS e MS/MS) a trappola ionica e q-TOF
• Synthesis and structural characterization, by mean of gas (GC-MS) and liquid chromatography (HPLC-ESI-MS) coupled to mass spectrometry, of farmacological compounds.
• Synthesis, structural characterization and biological application of aminoarylthiazoles for the correction of chloride transport in cystic fibrosis
• Synthesis, purification and structural characterization and quality control of creatine derivatives for the study of its transporter "in vivo" and for the therapy of its deficit.

Research group ERC sector

LS1_1, LS1_2, LS3_2, LS3_5, LS3_7, LS3_8, LS3_12, LS4_5, LS5_11, PE5_23, PE5_16, PE4_3, PE4_6

Internal personnel i

Collaborations

International collaborations:

• Prof. HC Lee - Peking University Shenzhen Graduate School, Pechino, Cina
• Prof. AH Guse - University Medical Center Hamburg-Eppendorf, Amburgo, Germania
• Prof. L Lamattina - Universidad Nacional de Mar del Plata, Mar del Plata, Argentina
• Prof.ssa I Robina - Department of Organic Chemistry, University of Seville, Spagna
• Prof. P Vogel - Laboratory of Glycochemistry and Asymmetric Synthesis, EPFL, Lausanne, Svizzera
• Prof. MW Sereda - Max-Planck-Institute for Experimental Medicine, Göttingen, Germania
• Prof. C.l Abergel - Institut de Microbiologie de la Méditerranée, Parc Scientifique de Luminy, Marseille, Francia
• Dr. S. Haslam - Department of Life Sciences, Imperial College Londra
• Prof. J. Van Etten - University of Nebraska-Lincoln, Department of Plant Pathology, Morrison Center Lincoln, Nebraska
• Dr. T. Hennet - Zurich Center for Integrative Human Physiology (ZIHP), Zurigo

National collaborations:

• Prof. M Magnani - Department of Biomolecular Sciences, Università of Urbino
• Prof. M Balestrino - Department of Neuroscience, Ophthalmology and Genetics, University of Genova
• Dott. P Ameri - Department of Internal Medicine, University of Genova
• Prof. G Murialdo - Department of Internal Medicine, University of Genova
• Prof. F Beguinot - Dept. of Cell and Molecular Biology and pathology, Univ of Napoli Federico II, Napoli
• Prof. A Nencioni - Department of Internal Medicine, University of Genova
• Prof. M Rizzi - Dipartimento di Scienze Chimiche, Alimentari, Farmaceutiche e Farmacologiche, Università del Piemonte Orientale Amedeo Avogadro, Novara
• Prof.ssa S Deaglio - Human Genetics Foundation e Università di Torino, Torino
• Prof.ssa N Raffaelli - Dipartimento di Scienze Agrarie, Alimentari ed Ambientali, Università Politecnica delle Marche, Ancona
• Prof. A Schenone - Dipartimento di Neuroscienze, Oftalmologia e Genetica, Università di Genova)