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Analytical Biochemistry and Proteomics Unit

Joint Unit Instituto de Investigaciones Biológicas Clemente Estable/Institut Pasteur de Montevideo

 

MEMBERS

  • Rosario Durán, PhD (Interim Head, Investigator IIBCE – IP Montevideo)
  • Carlos Batthyány, MD, PhD (Investigator-IP Montevideo; Adjunct Professor of Biochemistry, School of Medicine, UdelaR)
  • Magdalena Portela, BSc (Technical Assistant – School of Sciences/IP Montevideo)
  • Analía Lima, MSc. (Technical Assistant, PhD student)
  • Jessica Rossello, MSc. (Technician, PhD student; ANII Fellow)
  • Bernardina Rivera, BC, BSc.  (Technician, MSc. student; ANII fellow)
  • Alejandro Leyva BSc. (Technician, PhD student; ANII Fellow)
  • Jorge Rodríguez BSc. (PhD student; ANII Fellow)
  • Rosina Dapueto MSc (PhD student; ANII Fellow)
  • Germán Galliussi BSc. (MSc. student)
  • Rosina Toledo BSc. (MSc. student; ANII Fellow)

Associate Members

  • María Noel Álvarez, PhD (Associate Investigator, Adjunct Professor of Biochemistry, School of Medicine, UdelaR, Uruguay)
  • Andrés Kamaid, PhD (Associate Investigator)
  • Leonel Malacrida, PhD (Associate Investigator, Assistant Profesor, Pathophysiology Department, School of Medicine, UdelaR, Uruguay)
  • Virginia López, PhD (Adjunct Professor of Organic Chemistry, School of Chemistry and Science, UdelaR)

 

MAIN EQUIPMENT

  • HPLC, Agilent 1200
  • Capilar HPLC, Agilent 1200;
  • Nano HPLC, Easy-nLC 1000, Thermo
  • Nano HPLC Ultimate 300, Thermo
  • 2D Electrophoresis, EttanIPGphor + EttanDaltSix
  • Typhoon FLA 9500, GE Healthcare
  • 4800 MALDI TOF/TOF Mass Spectrometer, Abi Sciex
  • LTQ Velos + ETD Mass Spectrometer, Thermo
  • Q-exactive (Q-Orbitrap), Thermo

4800 MALDI TOF/TOF Mass Spectrometer, Abi Sciex

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LTQ Velos + ETD Mass Spectrometer, Thermo

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Q-exactive (Q-Orbitrap), Thermo

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SERVICES

Our Unit received in January 2007 a MALDI TOF-TOF MS instrument (AB-SCIEX, Framingham, USA) and, in December 2009, completed the MS platform with the arrival of a nano-electrospray/ion trap LTQ Velos instrument (Thermo, USA). Both instruments complement each other and expand the quality and type of mass analytical procedures we can offer to local and regional research groups.

There are two general modes to get access to the Unit facilities:

Routine Service

For routine analysis, users are welcome to access the UByPA as a “fee for service facility” supported by the Institut Pasteur de Montevideo. The facility offers this kind of service to researchers in the region, with priority given to users from the Institute and local academy. The analysis will be performed by members of our technical staff and will be done following standard protocols. The routine analysis includes analysis and interpretation of raw data based on routine practices only.

Routine analysis includes:

  • 2-D gel electrophoresis.
  • Protein sample preparation for MS analysis: in-gel digestion, in-solution digestion, desalting.
  • Molecular mass determination for peptides and small proteins by MS.
  • Protein identification by MALDI-TOF/TOF MS (peptide mass fingerprinting, MS/MS ion search) and database search.

In 2013 we analyzed 1178 different samples from 83 different groups: 55% of the samples were from our local academy, 42% were from the regional academy (Argentine, Brazil, Chile, Colombia, and Venezuela) and 3% from private industry.

In 2014 we analyzed 1238 samples from different groups from local and regional academy.

Non-Routine Service

Collaborative research projects, beyond routine services, are welcome. Members of the Unit are expected to significantly contribute to the conception, design of experiments and custom-design protocols, original ideas as well as data analysis and interpretation beyond routine practice.

Non routine analysis includes:

  • Custom sample preparation.
  • Post-translational modification analysis.
  • 2-D gel electrophoresis based proteomics.
  • “Shotgun” based proteomics.
  • Quantitative proteomics.
  • De novo peptide sequencing.
  • Glycomics and glycoproteomics.

Forms:

  1. Determinación de masa molecular de péptidos y proteínas pequeñas por MS
  2. Electroforesis Bidimensional
  3. Identifcación de proteínas por MALDI-TOF MS/MS

PRICES

Prices for routine services (american dollar, USD)

Academic Nacional Academic Regional* Academic International/Non-academic users
Molecular mass determination 12 36 72
2D Gel 10*10 cm + staining 60 240 480
2D Gel 20*20 cm + staining ask ask ask
2D Clean Up Kit for protein purification 12 48 96
2D Quant Kit protein quantitation 8 36 72
Protein identification by MALDI-TOF/TOF MS 24 60 165
  • *Latin America

 

RESEARCH

In the past years, members of our group have been involved in different areas of biological/biochemical research. A major contribution made by the UByPA´s scientist was the incorporation of modern mass spectrometry (MS) and 2D-DIGE to our local academy, bringing totally new analytical capabilities to perform comprehensive protein studies, including posttranslational modifications of proteins and the ability to decode cell signaling networks.Nowadays, we are involved in three main areas of research concerning protein-mediated cell signaling events:

CELL SIGNALING PATHWAYS IN PATHOGENIC BACTERIA: A PROTEOMIC APPROACH

CELL SIGNALING PATHWAYS IN MYCOBACTERIA: A PROTEOMIC APPROACH

The ability of M. tuberculosis to inhibit phagosome maturation and to survive in the intracellular environment of host is central to tuberculosis pathogenicity. The signaling cascades mediated by Ser/Thr kinases are key players in these processes. In particular one of these enzymes, PknG, became of special interest as it was found to play dual roles in mycobacterial metabolism and pathogenesis through mechanisms still not completely understood.  In order to unveil PknG partners in mycobacteria we developed an affinity purification-sequential elution-mass spectrometry strategy to identify substrates and interactors of the kinase.  This strategy allowed us to identify new PknG substrates that were further validated by in vitro and aslo in vivo by proteomic analysis of a pknG null mutant strain. We provide evidence that PknG regulates nitrogen assimilation and cell wall synthesis in mycobacteria. These are essential processes not only for bacterial physiology but also for the pathogen survival in the nutrient deficient environment within the host. Interestingly, some of the identified substrates contain a Forkhead-associated domain (FHA).  The FHA domain is a protein module that specifically recognizes phospho-Thr residues and that participates in the assembly of multicomponent signaling complexes in transduction pathways regulated by phosphorylation in prokaryotes and eukaryotes. The genome of M. tuberculosis codifies five proteins with FHA domain, which have been reported as in vitro substrates of several Ser/Thr kinases. The presence of signaling networks between Ser/Thr kinases and FHA domain-containing proteins in mycobacteria has been postulated, however nowadays our knowledge about its architecture, the protein-protein interactions underlying this network and the biological process under control is very scarce. Our research aims to contribute to the elucidation of FHA domain mediated signaling networks in mycobacteria by  characterizing at the molecular level the signaling complex formed in vivo by FHA domain-containing proteins and its dynamics in response to the biochemical stimuli found in the host. We are focusing in two FHA domain-containing proteins, GarA and FhaA, which we identify as substrates of PknG. The experimental strategy proposed combines the specific purification of tagged proteins with in vivo crosslinking to provide a snapshot of protein interactions in the mycobacteria. This approach will contribute to unravel the processes controlled by PknG signaling cascades mediated by its FHA domain containing substrates and its possible role in the adaptation to the host environment.

BIOLOGICAL EFFECTS OF SECOND GENERATION NITROALKENE TOCOPHEROL ANALOGS:

Atherosclerosis is an epidemic worldwide disease and leading cause of death in developed countries. Two main pathogenic events are well recognized in the generation of the atheroma plaques: inflammation and lipid accumulation via LDL deposition/foam cells formation. Inflammation is a key event mediated by inflammasome activation by sterile signals (i.e. cholesterol crystals) and precedes massive lipid accumulation.

In our work we envision a new pharmacological strategy for the treatment and prevention of atherosclerosis. We designed a hybrid compound analog of -tocopherol to which we added the electrophilic nitroalkene group (1). The rationale for our idea is that the nitroalkene-tocopherol analog will be selectively incorporated into the lipoprotein particles during their normal metabolism due to the presence of the chromanol structure. Once incorporated, lipoproteins will transport the compound through the body, including to the atherosclerotic lesions, were it may exert the potent anti-inflammatory and anti-atherogenic properties of the nitroalkenes.

We synthesized two series of tocopherol analogs and performed their physicochemical and biological characterization. Nitroalkene-tocopherol analogs are electrophiles that are being incorporated into lipoproteins both in vitro and in vivo and exhibit potent anti-inflammatory activity: A- inhibit pro-inflammatory cytokines (IL-6, MCP-1, TNF) secretion controlled by NFB factor & B- induce expression of phase 2 enzymes regulated by Nrf2/Keap-1 (HO-1, GCLM, NQO1) in murine macrophages; C- inhibit interleukin 1- secretion by NLRP3 inflammasome in THP-1 cells. We are now evaluating their capabilities to inhibit the development of atherosclerotic plaques formation in two animal models: apo E-/- and zebrafish.

    1. Design and Development of Nitroalkene Tocopherol and Analogs for Use in the Treatment and Prevention of Inflammatory Mediated Diseases. PIs: C. Batthyány and V. López; Graduate Thesis Project of J. Rodriguez, PEDECIBA, UdelaR, 2012 to date).
    2. Electrophilic mediated protein modifications.
      1. “Composition and Method for Inhibition of PknG from Mycobacterium tuberculosis”, PIs C. Batthyány & R. Durán (U.S. PCT Application No. 61/835,416; 2014).
      2. Molecular Mechanisms of Nitroalkene mediated anti-inflammatory cell signaling events; PIs C. Batthyány & H. Botti.

 

EDUCATION-COURSES

ORGANIZATION OF COURSES

UNU-BIOLAC & PEDECIBA “Proteome Analysis by Mass Spectrometry”; November 28 to December 2, 2016. Organizers: Rosario Durán, Paulo Carvalho & Carlos Batthyány.
• UNU-BIOLAC & RIIP (Institut Pasteur International Network) “Proteome Analysis by Mass Spectrometry”;. September 1 to 12, 2014. Organizers: Rosario Durán & Carlos Batthyány.
• UNU-Biolac- “Mass Spectrometry (MS) in Proteomics” Institut Pasteur de Montevideo – 26 November-8 December, 2012.
• UNU-Biolac – “Métodos Básicos en Proteómica”, Universidad Nacional de Asunción, Asunción del Paraguay, Paraguay, 2010.
• EMBO – “International Practical Course in Mass Spectrometry”, Institut Pasteur de Montevideo, Uruguay, 2010.
• Universidad de Sao Paulo – “Taller sobre La Espectrometría de Masa en la Proteómica”, Facultad de Ciencias Farmacéuticas de Ribeirão Preto, Universidad de São Paulo, Ribeirao Preto, Brasil, 2009.

GRANTS

  1. “Development of a novel class of anti-atherogenic agents: electrophilic nitroalkenes-Vitamin E (α-tocopherol) analogs”. (2013 – 2015); CABBIO; PI Carlos Batthyány; Amount Granted USD 30.000.
  2. “N‐Glycan fingerprint of exosomes in Chagas and Cancer diseases”. (2014); Interdisciplinary projects IP Montevideo; Coordinator C. Batthyány, Amount granted USD 20.000.
  3. Exploring the role of mosquito’s saliva in the transmission of Rift Valley fever” (2012-2014); Actions Concertées Interpasteuriennes (ACIP). Scientific coordinator: V. CHOUMET (Paris). Uruguayan PIs: C. Batthyány & R. Durán. Amount Granted: EUR 18.000.
  4. “Caracterización nutricional y de compuestos bioactivos del trigo en Uruguay. Variabilidad de genotipos y ambientes”; (2014 – 2016); FPTA – INIA: Contrato de Servicio PIs C. Batthyány & L. Malacrida; Amount Granted USD 32.000.
  5. “Identification of tumor associated antigens”; (2014 – 2015); Private Company: Contrato de Servicio; PIs R. Durán & C. Batthyány; Amount Granted: USD 35.500.
  6. Anti-atherogenic effects and molecular mechanisms of nitroalkene tocopherol analgos: a novel pharmacological approach” (2014-2016); PIs C. Batthyány & H. Botti; Amount Granted: USD 30.000.
  7. Análisis proteómico comparativo de dos cepas de P. aeruginosa con distinta capacidad de adhesión a células epiteliales” (2014-2016); J. Rossello (FCE_3_2013_1_100344); Amount Granted: USD 25.000.
  8. Hacia la elucidación del mecanismo molecular utilizado por PknG para ejercer su rol como factor de virulencia” (2014-2016). M. Gil (FCE_3_2013_1_100358); Amount Granted: USD 20.000.
  9. Surfactante Pulmonar durante la Lesión Pulmonar Aguda: Abordaje estructural, dinámico y funcional (2013 – 2015) CSIC I+D 2012, PI: L. Malacrida, Amount Granted: USD 38.000.
  10. “Redes de señalización mediadas por dominios FHA en micobacterias y su rol en la adaptación al ambiente del hospedero” (2015-2018); R. Durán (FCE_1_2014_1_104045); Amount Granted: USD 50.000
  11. Desarrollo y validación de procesos para el estudio y valorización de nutracéuticos: creación de la primer empresa uruguaya del tipo “Contract of Research Organization”. (2016-2018); C. Batthyany (ALI_2-2014-1-5055); Amount Granted: USD 302.000
  12. Development, synthesis and characterization of novel anti-inflammatory compounds”. (2016-2018); C. Batthyany, V. López, C. Escande (CITES, http://cites-gss.com/); Amount Granted: USD 630.000.

PUBLICATIONS

PATENTS

  1. “Methods of treatment of inflammation related conditions using pluripotent anti-inflammatory and metabolic modulators”; inventors Batthyany, C., Lopez, G.V., Escande, C., Porcal, W., Dapueto, R., Rodriguez, R., Galliussi, G., and Garat, M.P. 2016. USA patent provisional application; to be assigned.
  2. “Trolox derivatives and methods of use thereof in the treatment and prevention of inflammation related conditions”; inventors Batthyany, C., Lopez, G.V., Dapueto, R., Escande, C., and Rodriguez, R. 2016. USA patent non-provisional application; to be assigned.
  3. “System, Method and Device for Identifying Discriminant Biological Factors and for Classifying proteomic profiles”; inventors Carvalho PC, Batthyany C, Silva A; Lima D; Leyva A; Barbaosa V; Durán, R. 2016, USA provisional patent application; to be assigned
  4. “Tocopherol and Analogs for Use in the Treatment and Prevention of Inflammation Related Conditions”; (U.S. PCT Application WO 2015/073527 A1; 2015; co-inventores C.Batthyany & G.V.López).
  5. “Composition and Method for Inhibition of PknG from Mycobacterium Tuberculosis”; (U.S. PCT Application WO 2014; co-inventores C.Batthyany & R. Durán).

 

ARTICLES

 

  • Cabrera G, Lundberg U, Rodríguez-Ulloa A, Herrera M, Machado W, Portela M, Palomares S, Espinosa LA, Ramos Y, Durán R, Besada V, Vonasek E, González LJ  Protein content of the Hylesia metabus egg nest setae (Cramer [1775]) (Lepidoptera: Saturniidae) and its association with the parental investment for the reproductive success and lepidopterism. J Proteomics 150:183-200 (2017).
  • Batthyány C, Bartesaghi S, Mastrogiovanni M, Lima A, Demicheli V, Radi R.
    Antioxid Redox Signal. 2017 Mar 1;26(7):313-328. doi: 10.1089/ars.2016.6787. Epub 2016 Jul 22.
    Tyrosine-Nitrated Proteins: Proteomic and Bioanalytical Aspects.
  • Folle AM, Kitano ES, Lima A, Gil M, Cucher M, Mourglia-Ettlin G, Iwai LK, Rosenzvit M, Batthyány C, Ferreira AM.PLoS Negl Trop Dis. 2017 Jan 3;11(1):e0005250. doi: 10.1371/journal.pntd.0005250. eCollection 2017.
    Characterisation of Antigen B Protein Species Present in the Hydatid Cyst Fluid of Echinococcus canadensis G7 Genotype.
  • Silva AR, Lima DB, Peña A, Duran R, Batthyany C, Aquino PF, Leal JC, Rodriguez JE, Domont GB, Santos MD, Chamot-Rooke J, Barbosa VC, Carvalho PC  DiagnoProt: a tool for discovery of new molecules by mass spectrometry. Bioinformatics. Feb 10. doi: 10.1093/bioinformatics/btx093 (2017).
  • Fló M, Margenat M, Pellizza L, Graña M, Durán R, Báez A, Salceda E, Soto E, Alvarez B, Fernández C.Functional diversity of secreted cestode Kunitz proteins: Inhibition of serine peptidases and blockade of cation channels. PLoS Pathog. 13(2):e1006169 (2017) .
  •  Demicheli, D.M. Moreno, G.E . Jara ,A. Lima, S. Carballal , N. Ríos, C. Batthyany , G  Ferrer-Sueta , C.  Quijano ,  D.A. Estrı́n,  M.A. Martí, R.  Radi. Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34. Biochemistry. 55 (2016) 3403-3417.
  • E. Dieterle, J. Fina Martin, R. Durán, S.I. Nemirovsky, C. Sanchez Rivas, C. Bowman, D. Russell, G.F. Hatfull, C. Cambillau, M. Piuri. Characterization of prophages containing “evolved” Dit/Tal modules in the genome of Lactobacillus casei BL23, Applied Microbiology and Biotechnology 100 (2016) 9201-9215.
  • Cabrera, U. Lundberg, A. Rodriguez-Ulloa, M. Herrera, W. Machado, M. Portela, S. Palomares, L.A. Espinosa, Y. Ramos, R. Duran, V. Besada, E. Vonasek, L.J. Gonzalez. , Protein content of the Hylesiametabus egg nest setae (Cramer [1775]) (Lepidoptera: Saturniidae) and its association with the parental investment for the reproductive success and lepidopterism, J Proteomics 150 (2016) 183-200.
  • Wagner, T., Alexandre, M., Durán, R., Barilone, N., Wehenkel, A., Alzari, P.M.& Bellinzoni, M.. The crystal structure of the catalytic domain of the ser/thr kinase PknA from tuberculosis shows an Src-like autoinhibited conformation. Proteins 83:982-988.(2015).
  • Lisa, M.N., Gil, M., André-Leroux, G., Barilone, N., Durán, R., Biondi, R.M. & Alzari, P.M.. Molecular Basis of the Activity and the Regulation of the Eukaryotic-like S/T Protein Kinase PknG from Mycobacterium tuberculosis. Structure 23:1039-1048. (2015).
  • Margenat ,M., Labandera, A.M., Gil, M., Carrion, F., Purificação, M., Razzera, G., Portela, M.M., Obal, G., Terenzi, H., Pritsch, O., Durán, R., Ferreira, A.M. & Villarino, A. New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state. Sci. Rep.5: 8819. (2015).
  • Yunes Quartino, P.J., Portela, M., Lima, A., Durán, R., Lomonte, B. & Fidelio, G.D. 2015. A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipase A2. Biochim Biophys Acta.1848: 216-224.
  • Randall, L. M., Manta, B., Hugo, M., Gil, M., Batthyány, C., Trujillo, M., Poole, L. B., and Denicola, A. Nitration transforms a sensitive peroxiredoxin 2 into a more active and robust peroxidase, The Journal of biological chemistry 289, 15536-15543. (2014).
  • Dieterle, M. E., Bowman, C., Batthyány, C., Lanzarotti, E., Turjanski, A., Hatfull, G., and Piuri, M. (2014) Exposing the secrets of two well known Lactobacillus casei phages: Genomic and structural analysis of J-1 and PL-1, Applied and environmental microbiology. 80(22):7107-21.
  • Mon, M.L., Moyano, R.D., Viale, M.N., Colombatti Olivieri, M.A., Gamietea, I.J., Montenegro, V.N., Alonso, B., Santangelo, M.D.L.P., Singh, M., Durán, R., Romano, M.I. Evaluation of cocktails with recombinant proteins of mycobacterium bovis for a specific diagnosis of bovine tuberculosis BioMed Research International, 2014, art. no. 140829. (2014).
  • Martinez, A., Peluffo, G., Petruk, A.A., Hugo, M., Piñeyro, D., Demicheli, V., Moreno, D.M., Lima, A., Batthyány, C., Durán, R., Robello, C., Martí, M.A., Larrieux, N., Buschiazzo, A., Trujillo, M., Radi, R., Piacenza, L. Structural and molecular basis of the peroxynitrite-mediated nitration and inactivation of Trypanosoma cruzi iron-superoxide dismutases (Fe-SODs) A and B: Disparate susceptibilities due to the repair of Tyr35 radical by Cys83 in Fe-SODB through intramolecular electron transfer. Journal of Biological Chemistry, 289: 12760-12778. (2014).
  • Alvarez G, Aguirre-López B, Cabrera N, Marins EB, Tinoco L, Batthyány CI, de Gómez-Puyou MT, Puyou AG, Pérez-Montfort R, Cerecetto H, González M. 1,2,4-thiadiazol-5(4H)-ones: a new class of selective inhibitors of Trypanosoma cruzi triosephosphate isomerase. Study of the mechanism of inhibition. J Enzyme Inhib Med Chem. 28:981-9 (2013).
  • Gil, M., Graña, M., Schopfer, F. J., Wagner, T., Denicola, A., Freeman, B. A., Alzari, P. M., Batthyány, C., and Durán, R. Inhibition of Mycobacterium tuberculosis PknG by non-catalytic rubredoxin domain specific modification: reaction of an electrophilic nitro-fatty acid with the Fe-S center, Free Radical Biology & Medicine 65: 150-161. (2013).
  • Basika T; Muñoz N; Casaravilla C; Irigoin F; Batthyany, C.; Bonilla M; Salinas, G; Pacheco JP; Roth J; Durán, R; Díaz A. (2012). Phagocyte-specific S100 proteins in the local response to the Echinococcus granulosus larva. Parasitology, 139: 271-283.
  • Obal, G.; Ramos, A. L.; Silva, V.; Lima, A.; Batthyány, C.; Bessio, M.L.; Ferreira, F.; Salinas, G.; Ferreira, A. (2012). Characterisation of the native lipid moiety of Echinococcus granulosus antigen B. PLoS Neglected Tropical Diseases, v.: 6 5.
  • Bonacci G, Baker PR, Salvatore SR, Shores D, Khoo NK, Koenitzer JR, Vitturi DA, Woodcock SR, Golin-Bisello F, Cole MP, Watkins S, St Croix C, Batthyany CI, Freeman BA, Schopfer FJ. (2012). Conjugated linoleic acid is a preferential substrate for fatty acid nitration. J Biol Chem. 287(53):44071-82.
  • Laschuk A, Monteiro KM, Vidal NM, Pinto PM, Duran R, Cerveñansky C, Zaha A, Ferreira HB. Proteomic survey of the cestode Mesocestoides corti during the first 24 hours of strobilar development. Parasitol Res. 108, 645-56 (2011).
  • Binolfi A, Valiente-Gabioud AA, Durán R, Zweckstetter M, Griesinger C, Fernandez CO Exploring the Structural Details of Cu(I) Binding to α-Synuclein by NMR Spectroscopy. Am. Chem. Soc., 133 :194–196 (2011).
  • Bonacci G, Schopfer FJ, Batthyany CI, et al. Electrophilic Fatty acids regulate matrix metalloproteinase activity and expression. J Biol Chem  286:16074-81(2011).
  • Lima A, Durán R, Schujman G, Marchissio MJ, Portela MM, Obal G, Pritsch O, de Mendoza D, Cerveñansky C. Serine/threonine protein kinase PrkA of the human pathogen Listeria monocytogenes: Biochemical characterization and identification of interacting partners through proteomic approaches. J Proteomics 74:1720-34 (2011).
  • Garavaglia PA, Cannata JJ, Ruiza AM, Maugeric D, Durán R, Galleanoe M,  García GA. Identification, cloning and characterization of an aldo-keto reductase from Trypanosoma cruzi with quinone oxidoreductase activity. Mol Biochem Parasitol. 173, 131-141 (2010).
  • Binolfi A, Rodriguez EE, Valensin D, D’Amelio N, Ippoliti E, Obal G, Durán R, Magistrato A, Pritsch O, Zweckstetter M, Valensin G, Carloni P, Quintanar L, Griesinger C, Fernández CO.  Bioinorganic chemistry of Parkinson’s disease: structural determinants for the copper-mediated amyloid formation of alpha-synuclein.  Inorg Chem. 49:10668-79 (2010).
  • Schopfer FJ, Cole MP, Groeger AL, Chen CS, Khoo NK, Woodcock SR, Golin-Bisello F, Motanya UN, Li Y, Zhang J, Garcia-Barrio MT, Rudolph TK, Rudolph V, Bonacci G, Baker PR, Xu HE, Batthyany CI, Chen YE, Hallis TM, Freeman BA. Covalent peroxisome proliferator-activated receptor gamma adduction by nitro-fatty acids: selective ligand activity and anti-diabetic signaling actions. J Biol Chem 285:12321-33 (2010).
  • Khoo NK, Rudolph V, Cole MP, Golin-Bisello F, Schopfer FJ, Woodcock SR, Batthyany C, Freeman BA. Activation of vascular endothelial nitric oxide synthase and heme oxygenase-1 expression by electrophilic nitro-fatty acids. Free Radic Biol Med 48:230-9 (2010).
  • Palacios F, Cota G, Horjales S, Lima A, Battistoni J, Sotelo-Silveira J, Marín M. An antibody-based affinity chromatography tool to assess Cu,Zn superoxide dismutase (SOD) G93A structural complexity in vivo. Biotechnol J. 5:328-34 (2010).
  • Celano L; Gil M,  Carballal S; Durán, R; Denicola A; Banerjeef R; Alvarez, B. nactivation of cystathionine;-synthase with peroxynitrite. Arch Biochem Biophys. 491, 1-2 (2009).
  • González S; Fló M; Margenat M; Durán, R; Gonzalez G; Graña, M.; Parkinson J; Maizels RM; Salinas G; Alvarez, B, Fernández C.A Family of Diverse Kunitz Inhibitors from Echinococcus granulosus Potentially Involved in Host-Parasite Cross-Talk PLoS ONE17, 4(9):e7009. (2009).
  • Sánchez G, Colettis N, Vázquez P, Cerveñansky C, Aguirre A, Quillfeldt JA, Jerusalinsky D, Kornisiuk E Muscarinic inhibition of hippocampal and striatal adenylyl cyclase is mainly due to the M(4) receptor. Neurochem Res. 34:1363-71 (2009).
  • Sánchez G, Alvares Lde O, Oberholzer MV, Genro B, Quillfeldt J, da Costa JC, Cerveñansky C, Jerusalinsky D, Kornisiuk E. M4 muscarinic receptors are involved in modulation of neurotransmission at synapses of Schaffer collaterals on CA1 hippocampal neurons in rats J Neurosci Res.87:691-700 (2009).
  • Díaz A, Fontana EC, Todeschini AR, Soulé S, González H, Casaravilla C, Portela M, Mohana-Borges R, Mendonça-Previato L, Previato JO, Ferreira F. The major surface carbohydrates of the Echinococcus granulosus cyst: mucin-type O-glycans decorated by novel galactose-based structures. Biochemistry15;48(49):11678-91 (2009).
  • Schopfer FJ, Batthyany C, Baker PR, et al. Detection and quantification of protein adduction by electrophilic fatty acids: mitochondrial generation of fatty acid nitroalkene derivatives. Free Radic Biol Med  46:1250-9 (2009).
  • Ferreira AM, Ferrari MI, Trostchansky A, Batthyany C, Souza JM, Alvarez MN, López GV, Baker PR, Schopfer FJ, O’Donnell V, Freeman BA, Rubbo H. Macrophage activation induces formation of the anti-inflammatory lipid cholesteryl-nitrolinoleate. Biochem J  417:223-34 (2009).
  • Wehenkel A, Bellinzoni M, Graña M, Durán R, Villarino A, Fernandez P, Andre-Leroux G, England P, Takiff H, Cerveñansky C, Cole ST, Alzari PM. Mycobacterial Ser/Thr protein kinases and phosphatases: physiological roles and therapeutic potential. Biochim Biophys Acta. 1784, 193-202 (2008).
  • Turell L, Botti H, Carballal S, Ferrer-Sueta G, Souza JM, Durán R, Freeman BA, Radi R, Alvarez B. Reactivity of sulfenic acid in human serum albumin.. Biochemistry 47, 358-367 (2008).
  • Irañeta S G, Acosta DM,  Durán R, Apicella C, Orlando UD,  Seoane MA, Alonso A and  Duschak VG. MALDI-TOF MS analysis of labile Lolium perenne major allergens in mixes.  Clin Exp Allergy 38, 1391-1399 (2008).
  • Cancela M., Acosta D., Rinaldi G, Silva, E., Durán, R., Roche L., Zaha, A., Carmona, C., Tort, J.F. A distinctive repertoire of cathepsins is expressed by juvenile invasive fasciola hepatica. Biochimie 90,1461-1475 (2008).
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CONTACT

ubypa@pasteur.edu.uy