1. Protein oxidation and carbonylation: proteomic and mass spectrometric approaches.
Our first interset is the identification and characterization of new pharmacological targets and biomarkers of oxidative/carbonyl stress associated to a variety of pathological conditions. The research is focussed on the development and application of mass spectrometric and immunochemical techniques for the identification of post-translational modifications of proteins in different biological matrices as early biomarkers of oxidative/carbonyl damage.
2. Development of new detoxifying agents of cytotoxic reactive carbonyl species (RCS).
By considering the lipoxidation-derived reactive carbonyl species (RCS) as potential drug targets in preventing protein carbonylation and related cellular dysfunction, the research project (in collaboration with academic and industrial partners) focuses on the development of new RCS detoxifying agents, using the natural dipeptide carnosine as active scaffold. The project involves an in vitro screening phase devoted to the evaluation of the biological activity and metabolic stability of the developed compounds, followed by in vivo evaluation of the pharmacokinetic and pharmacodynamic profiles (in experimental animal models of carbonyl stress) of the lead compounds identified in the in vitro approach.
Instrumentation
nano flow UPLC system (DIONEX UltiMate® 3000 Nano LC systems) connected to the ORBITRAP MS analyzer
LC-MS/MS system LTQ Orbitrap XL™ hybrid FTMS (Fourier Transform Mass Spectrometer) with high resolution, mass accuracy and dynamic range trougth a nano ESI source
Semi-preparative HPLC system connected to an UV-DAD detector and an automatic fractions collector (VARIAN LC-940)
HPLC-UV-MS/MS system (Thermo Finnigan Quantum Ultra) equipped with an UV-DAD detector, a triple quadrupole mass spectrometer MS/MS with ESI (Electrospray) and APCI (Atmosheric Pressure Ionization) interfaces, and a Xcalibur® system manager data acquisition software
HPLC-UV-MS/MS system (Thermo Finnigan Advantage) equipped with an UV-DAD detector, an Ion Trap mass spectrometer with ESI (Electrospray), nano-ESI and APCI (Atmosheric Pressure Ionization) interfaces, and a Xcalibur system manager data acquisition software
Techniques
Mass spectrometric techniques for protein identification and characterization of oxidized/carbonylated sites. Design and evaluation of novel RCS sequestering agents. Metabolic pre-screening and ADMET studies.
Selected references
1: Aldini G, Orioli M, Rossoni G, Savi F, Braidotti P, Vistoli G, Yeum KJ, Negrisoli G, Carini M. The carbonyl scavenger carnosine ameliorates dyslipidemia and renal function in zucker obese rats. J Cell Mol Med. 2010 Jun 1. [Epub aheadof print] PubMed PMID: 20518851.
2: Vistoli G, Orioli M, Pedretti A, Regazzoni L, Canevotti R, Negrisoli G, Carini M, Aldini G. Design, synthesis, and evaluation of carnosine derivatives as selective and efficient sequestering agents of cytotoxic reactive carbonyl species. ChemMedChem. 2009 Jun;4(6):967-75. PubMed PMID: 19301317.
3: Aldini G, Regazzoni L, Orioli M, Rimoldi I, Facino RM, Carini M. A tandem MS precursor-ion scan approach to identify variable covalent modification of albumin Cys34: a new tool for studying vascular carbonylation. J Mass Spectrom. 2008 Nov;43(11):1470-81. PubMed PMID: 18457351.
4: Aldini G, Vistoli G, Regazzoni L, Gamberoni L, Facino RM, Yamaguchi S, Uchida K, Carini M. Albumin is the main nucleophilic target of human plasma: a protective role against pro-atherogenic electrophilic reactive carbonyl species? Chem Res Toxicol. 2008 Apr;21(4):824-35. Epub 2008 Mar 7. PubMed PMID: 18324789.
5: Orioli M, Aldini G, Benfatto MC, Facino RM, Carini M. HNE Michael adducts to histidine and histidine-containing peptides as biomarkers of lipid-derived carbonyl stress in urines: LC-MS/MS profiling in Zucker obese rats. Anal Chem. 2007 Dec 1;79(23):9174-84. Epub 2007 Nov 3. PubMed PMID: 17979257.
6: Dalle-Donne I, Carini M, Vistoli G, Gamberoni L, Giustarini D, Colombo R, Maffei Facino R, Rossi R, Milzani A, Aldini G. Actin Cys374 as a nucleophilic target of alpha,beta-unsaturated aldehydes. Free Radic Biol Med. 2007 Mar 1;42(5):583-98. Epub 2006 Nov 30. PubMed PMID: 17291982.
7: Aldini G, Dalle-Donne I, Facino RM, Milzani A, Carini M. Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonyls. Med Res Rev. 2007 Nov;27(6):817-68. Review. PubMed PMID: 17044003.
8: Aldini G, Gamberoni L, Orioli M, Beretta G, Regazzoni L, Maffei Facino R, Carini M. Mass spectrometric characterization of covalent modification of human serum albumin by 4-hydroxy-trans-2-nonenal. J Mass Spectrom. 2006 Sep;41(9):1149-61. PubMed PMID: 16888752.
9: Aldini G, Dalle-Donne I, Vistoli G, Maffei Facino R, Carini M. Covalent modification of actin by 4-hydroxy-trans-2-nonenal (HNE): LC-ESI-MS/MS evidence for Cys374 Michael adduction. J Mass Spectrom. 2005 Jul;40(7):946-54. PubMed PMID: 15934040.
10: Aldini G, Carini M, Beretta G, Bradamante S, Facino RM. Carnosine is a quencher of 4-hydroxy-nonenal: through what mechanism of reaction? Biochem Biophys Res Commun. 2002 Nov 15;298(5):699-706. PubMed PMID: 12419310.
ALDINI Giancarlo
Contacts
Contacting person: Giancarlo Aldini; e-mail: giancarlo.aldini@unimi.it Full address: Deapertment of Pharmaceutical Sciences “Pietro Pratesi” – University of Milan via Mangiagalli 25, 20133 Milan, ItalyPhone number: +39-02-50319296
Fax number: +39-02-50319359 Skype: galdini67
Group members
Marina Carini, Giulio Vistoli, Marica Orioli, Luca Regazzoni
Research interests
Our first interset is the identification and characterization of new pharmacological targets and biomarkers of oxidative/carbonyl stress associated to a variety of pathological conditions. The research is focussed on the development and application of mass spectrometric and immunochemical techniques for the identification of post-translational modifications of proteins in different biological matrices as early biomarkers of oxidative/carbonyl damage.
2. Development of new detoxifying agents of cytotoxic reactive carbonyl species (RCS).
By considering the lipoxidation-derived reactive carbonyl species (RCS) as potential drug targets in preventing protein carbonylation and related cellular dysfunction, the research project (in collaboration with academic and industrial partners) focuses on the development of new RCS detoxifying agents, using the natural dipeptide carnosine as active scaffold. The project involves an in vitro screening phase devoted to the evaluation of the biological activity and metabolic stability of the developed compounds, followed by in vivo evaluation of the pharmacokinetic and pharmacodynamic profiles (in experimental animal models of carbonyl stress) of the lead compounds identified in the in vitro approach.
Instrumentation
Techniques
Mass spectrometric techniques for protein identification and characterization of oxidized/carbonylated sites. Design and evaluation of novel RCS sequestering agents. Metabolic pre-screening and ADMET studies.
Selected references
1: Aldini G, Orioli M, Rossoni G, Savi F, Braidotti P, Vistoli G, Yeum KJ, Negrisoli G, Carini M. The carbonyl scavenger carnosine ameliorates dyslipidemia and renal function in zucker obese rats. J Cell Mol Med. 2010 Jun 1. [Epub aheadof print] PubMed PMID: 20518851.
2: Vistoli G, Orioli M, Pedretti A, Regazzoni L, Canevotti R, Negrisoli G, Carini M, Aldini G. Design, synthesis, and evaluation of carnosine derivatives as selective and efficient sequestering agents of cytotoxic reactive carbonyl species. ChemMedChem. 2009 Jun;4(6):967-75. PubMed PMID: 19301317.
3: Aldini G, Regazzoni L, Orioli M, Rimoldi I, Facino RM, Carini M. A tandem MS precursor-ion scan approach to identify variable covalent modification of albumin Cys34: a new tool for studying vascular carbonylation. J Mass Spectrom. 2008 Nov;43(11):1470-81. PubMed PMID: 18457351.
4: Aldini G, Vistoli G, Regazzoni L, Gamberoni L, Facino RM, Yamaguchi S, Uchida K, Carini M. Albumin is the main nucleophilic target of human plasma: a protective role against pro-atherogenic electrophilic reactive carbonyl species? Chem Res Toxicol. 2008 Apr;21(4):824-35. Epub 2008 Mar 7. PubMed PMID: 18324789.
5: Orioli M, Aldini G, Benfatto MC, Facino RM, Carini M. HNE Michael adducts to histidine and histidine-containing peptides as biomarkers of lipid-derived carbonyl stress in urines: LC-MS/MS profiling in Zucker obese rats. Anal Chem. 2007 Dec 1;79(23):9174-84. Epub 2007 Nov 3. PubMed PMID: 17979257.
6: Dalle-Donne I, Carini M, Vistoli G, Gamberoni L, Giustarini D, Colombo R, Maffei Facino R, Rossi R, Milzani A, Aldini G. Actin Cys374 as a nucleophilic target of alpha,beta-unsaturated aldehydes. Free Radic Biol Med. 2007 Mar 1;42(5):583-98. Epub 2006 Nov 30. PubMed PMID: 17291982.
7: Aldini G, Dalle-Donne I, Facino RM, Milzani A, Carini M. Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonyls. Med Res Rev. 2007 Nov;27(6):817-68. Review. PubMed PMID: 17044003.
8: Aldini G, Gamberoni L, Orioli M, Beretta G, Regazzoni L, Maffei Facino R, Carini M. Mass spectrometric characterization of covalent modification of human serum albumin by 4-hydroxy-trans-2-nonenal. J Mass Spectrom. 2006 Sep;41(9):1149-61. PubMed PMID: 16888752.
9: Aldini G, Dalle-Donne I, Vistoli G, Maffei Facino R, Carini M. Covalent modification of actin by 4-hydroxy-trans-2-nonenal (HNE): LC-ESI-MS/MS evidence for Cys374 Michael adduction. J Mass Spectrom. 2005 Jul;40(7):946-54. PubMed PMID: 15934040.
10: Aldini G, Carini M, Beretta G, Bradamante S, Facino RM. Carnosine is a quencher of 4-hydroxy-nonenal: through what mechanism of reaction? Biochem Biophys Res Commun. 2002 Nov 15;298(5):699-706. PubMed PMID: 12419310.