Solange M T Serrano

Instituto Butantan, San Paulo, São Paulo, Brazil

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Publications (77)232.54 Total impact

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    ABSTRACT: Many snake venom toxins are serine proteases but their specific in vivo targets are mostly unknown. Various act on components of the coagulation cascade, and fibrinolytic and kallikrein-kinin systems to trigger various pathological effects observed in the envenomation. Despite showing high similarity in terms of primary structure snake venom serine proteinases (SVSPs) show exquisite specificity towards macromolecular substrates. Therefore, the characterization of their peptide bond specificity is important for understanding the active site preference associated with effective proteolysis as well as for the design of peptide substrates and inhibitors. Bothrops jararaca contains various SVSPs among which Bothrops protease A is a specific fibrinogenolytic agent and PA-BJ is a platelet-activating enzyme. In this study we used proteome derived peptide libraries in the Proteomic Identification of protease Cleavage Sites (PICS) approach to explore the peptide bond specificity of Bothrops protease A and PA-BJ in order to determine their individual peptide cleavage sequences. A total of 371 cleavage sites (208 for Bothrops protease A and 163 for PA-BJ) were detected and both proteinases displayed a clear preference for arginine at the P1 position. Moreover, the analysis of the specificity profiles of Bothrops protease A and PA-BJ revealed subtle differences in the preferences along P6-P6', despite a common yet unusual preference for Pro at P2. Taken together, these results map the subsite specificity of both SVSPs and shed light in the functional differences between these proteinases. Proteolysis is key to various pathological effects observed upon envenomation by viperid snakes. The use of the Proteomic Identification of protease Cleavage Sites (PICS) approach for the easy mapping of proteinase subsite preferences at both the prime- and non-prime sides concurrently gives rise to a fresh understanding of the interaction of the snake venom serine proteinases with peptide and macromolecular substrates and indicates that their hydrolytic activity is influenced by the amino acid sequences adjacent to the scissile bond. Copyright © 2014. Published by Elsevier B.V.
    Journal of Proteomics. 10/2014;
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    ABSTRACT: Accidents caused by scorpions represent a relevant public health issue in Brazil, being more recurring than incidents with snakes and spiders. The main species responsible for this situation is the yellow scorpion, Tityus serrulatus, due especially to the great frequency with which accidents occur and the potential of its venom to induce severe clinical manifestations, even death, mainly among children. Although neurotoxins are well characterized, little information is known about other components of scorpion venoms, such as peptidases, and their effect on envenomation. Previous results from our group showed that the metallopeptidases present in this venom are capable of hydrolyzing the neuropeptide dynorphin 1-13 in vitro, releasing Leu-enkephalin, which may interact with ion channels and promote indirect neurotoxicity. Thus, this study aims to get more information about the effect of toxic peptidase activity present in the venom on biologically active peptides, and to evaluate the in vitro neutralizing potential of commercial antivenoms produced by the Butantan Institute. A set of human bioactive peptides were studied as substrates for the peptidases, and the members of the neuropeptide Y family were found to be the most susceptible ones. All new substrate hydrolyses were totally inhibited by EDTA and not blocked by PMSF, indicating that metallopeptidases were responsible for the peptidase activity. Also, peptidase activities were only partially inhibited by therapeutic Brazilian scorpion antivenom (SAV) and arachnid antivenom (AAV). The dose-response inhibition by both antivenoms indicates that AAV neutralizes better than SAV at the used doses. These characterizations, unpublished until now, can contribute to the improvement of our knowledge about the venom and envenomation processes by T. serrulatus.
    Toxicological sciences : an official journal of the Society of Toxicology. 09/2014;
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    ABSTRACT: Snake venoms contain serine proteinases that are functionally similar to thrombin and specifically cleave fibrinogen to convert it into fibrin or activate platelets to aggregation. PA-BJ is a serine proteinase from Bothrops jararaca venom that promotes platelet aggregation and this effect is mediated by the G-coupled protein receptors PAR1 and PAR4. In this study we describe an improved procedure to obtain PA-BJ from B. jararaca venom that uses less chromatographic steps, and, interestingly, results in the isolation of eight proteoforms showing slightly different pIs and molecular masses due to variations in their glycosylation levels. The identity of the isolated PA-BJ forms (1-8) was confirmed by mass spectrometry, and they showed similar platelet-activating activity on washed platelet suspensions. N- and O-deglycosylation of PA-BJ 1-8 under denaturing conditions generated variable electrophoretic profiles and showed that some forms were resistant to complete deglycosylation. Furthermore, N- and O-deglycosylation under non-denaturing conditions also showed different electrophoretic profiles between the PA-BJ forms and caused partial loss of their ability to cleave a recombinant exodomain of PAR1 receptor. In parallel, three cDNAs encoding PA-BJ-like enzymes were identified by pyrosequencing of a B. jararaca venom gland library constructed with RNA from a single specimen. Taken together, our results suggest that PA-BJ occurs in the B. jararaca venom in multiple proteoforms displaying similar properties upon platelets regardless of their variable isoelectric points, molecular masses, carbohydrate moieties and susceptibility to the activity of glycosidases, and highlight that variability of specific venom components contributes to venom proteome complexity.
    Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics 09/2014; · 3.73 Impact Factor
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    ABSTRACT: Antivenoms manufactured by bioCSL Limited (Australia) and Instituto Clodomiro Picado (Costa Rica) against the venom of the taipan snakes (Oxyuranus scutellatus) from Australia and Papua New Guinea (PNG), respectively, were compared using antivenomics, an analytical approach that combines proteomics with immunoaffinity chromatography. Both antivenoms recognized all venom proteins present in venom from PNG O. scutellatus, although a pattern of partial recognition was observed for some components. In the case of the Australian O. scutellatus venom, both antivenoms immunorecognized the majority of the components, but the CSL antivenom showed a stronger pattern of immunoreactivity, which was revealed by the percentage of retained proteins in the immunoaffinity column. Antivenoms interacted with taipoxin in surface plasmon resonance. These observations on antivenomics agree with previous neutralization studies.
    The American journal of tropical medicine and hygiene 08/2014; · 2.53 Impact Factor
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    ABSTRACT: Few virulence factors have been identified for Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis. In this study, we quantitatively evaluated the protein composition of P. brasiliensis in the yeast phase using minimal and rich media to obtain a better understanding of its virulence and to gain new insights into pathogen adaptation strategies. This analysis was performed on two isolates of the Pb18 strain showing distinct infection profiles in B10.A mice. Using liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis, we identified and quantified 316 proteins in minimal medium, 29 of which were overexpressed in virulent Pb18. In rich medium, 29 out of 295 proteins were overexpressed in the virulent fungus. Three proteins were found up-regulated in both media, suggesting potential roles of these proteins in virulence regulation in P. brasiliensis. Moreover, genes up-regulated in virulent Pb18 showed increase in its expression after the recovery of virulence of attenuated Pb18. Proteins up-regulated in both isolates were grouped according to their functional categories. Virulent Pb18 undergoes metabolic reorganization and increased expression of proteins involved in fermentative respiration. This approach allowed us to identify potential virulence regulators and provided a foundation for achieving a molecular understanding of how Paracoccidioides modulates the host-pathogen interaction to its advantage.
    Journal of Proteome Research 08/2014; · 5.06 Impact Factor
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    ABSTRACT: Background / Purpose: The Center of Toxins, Immune-response and Cell Signaling (CeTICS) is an emerging center that studies biochemical, molecular, and cellular mechanisms of toxins that have therapeutic potential. That aims to understand the short-term and long-term behavior of biological systems based on analysis of signaling networks. This is an interdisciplinary project, coupled to the heterogeneous, high-throughput data produced by modern large scale techniques in genomics and proteomics, which implies the necessity of data organization and integration to carry out scientific knowledge. Based on the project goals, we are developing a platform to provide a uniform conceptual schema to minimize -omics data representation. As well as a suitable semantics for interdisciplinary research that allows quantitative and qualitative -omics integrative analyses and mathematical modeling of signaling networks. Main conclusion: The CeTICSdb platform, in its present conceptual database model, allows different integration approaches, construction and simulation of dynamic models based on high-throughput omics data. In addition, it also allows retrieval, storage, management, and integration of experimental and simulated results. Further development of CeTICSdb’ database structure and data integration tools is planned to reduce gaps between behavior of experimental models and virtual simulated models.
    ISMB 2014 Satellite Meeting - HiTSeq SIG: High Throughput Sequencing Algorithms & Applications; 07/2014
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    ABSTRACT: By catalyzing limited proteolysis or extensive degradation, proteolytic enzymes determine the fate of most proteins in an organism. In the evolutionary process of snake venoms, genes encoding proteinases were tailored to generate potent toxins to target key physiological proteins and thereby play a critical role in prey capture, immobilization and defense against predators. In Bothrops jararaca, metalloproteinases and serine proteinases are among the most abundant toxins both in newborn and adult venoms. In this review, we examine the proteinase-rich venom proteome of B. jararaca and how the proteinases act in a complex and heterogeneous fashion to exert their deleterious local and systemic effects.
    Toxin Reviews. 06/2014;
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    ABSTRACT: Background / Purpose: The Center of Toxins, Immune-response and Cell Signaling (CeTICS) is an emerging center that studies biochemical, molecular, and cellular mechanisms of toxins that have therapeutic potential, aiming to develop proofs of concept based on analyses of molecular signaling networks. Once those studies and analyses involve boundary crossing between Biology, Mathematics, Statistics, and Computer Science, the research developed in CeTICS is intrinsically interdisciplinary. This fact, coupled to the huge amount of data produced by modern high-throughput methods in genomics and proteomics, implies the necessity of data organization and integration to yield scientific knowledge. To this end, we are developing CeTICSdb, a platform to provide a uniform conceptual schema to minimize -omics data representation, as well as a suitable semantics for interdisciplinary research that allows quantitative and qualitative -omics integrative analyses and mathematical modeling of signaling networks. Main conclusion: The CeTICSdb conceptual database model supports high-throughput -omics data and also the management, access and integration of such results. However, further development of the database structure and data integration tools are required to reduce the gap between the experimental models and the actual data representation. Having those requirements properly addressed, a mid-term objective is to make the CeTICSdb platform available to the scientific community as a dry lab, in which high-throughput -omics data could be used to design and validate reaction kinetics models.
    18th International Conference on Research in Computational Molecular Biology (RECOMB) 2014; 05/2014
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    ABSTRACT: Snake venom metalloproteinases (SVMPs) play important roles in the local and systemic hemorrhage observed upon envenomation. In a previous study on the structural elements important for the activities of HF3 (highly hemorrhagic, P-III-SVMP), bothropasin (hemorrhagic, P-III-SVMP) and BJ-PI (non-hemorrhagic, P-I-SVMP), from Bothrops jararaca, it was demonstrated that they differ in their proteolysis profile of plasma and extracellular matrix proteins. In this study, we evaluated the ability of proteins DM43 and α2-macroglobulin to interfere with the proteolytic activity of these SVMPs on fibrinogen and collagen VI and with their ability to induce hemorrhage. DM43 inhibited the proteolytic activity of bothropasin and BJ-PI but not that of HF3, and was not cleaved the three proteinases. On the other hand, α2-macroglobulin did not inhibit any of the proteinases and was rather cleaved by them. In agreement with these findings, binding analysis showed interaction of bothropasin and BJ-PI but not HF3 to DM43 while none of the proteinases bound to α2-macroglobulin. Moreover, DM43 promoted partial inhibition of the hemorrhagic activity of bothropasin but not that of HF3. Our results demonstrate that metalloproteinases of B. jararaca venom showing different domain composition, glycosylation level and hemorrhagic potency show variable susceptibilities to protein inhibitors.
    Toxicon 01/2014; · 2.92 Impact Factor
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    ABSTRACT: Snake venom metalloproteinases (SVMPs) are major components in most viperid venoms that induce disturbances in the hemostatic system and tissues of animals envenomated by snakes. These disturbances are involved in human pathology of snake bites and appear to be essential for the capture and digestion of snake's prey and avoidance of predators. SVMPs are a versatile family of venom toxins acting on different hemostatic targets which are present in venoms in distinct structural forms. However, the reason why a large number of different SVMPs are expressed in some venoms is still unclear. In this study, we evaluated the interference of five isolated SVMPs in blood coagulation of humans, birds and small rodents. P-III class SVMPs (fractions Ic, IIb and IIc) possess gelatinolytic and hemorrhagic activities, and, of these, two also show fibrinolytic activity. P-I class SVMPs (fractions IVa and IVb) are only fibrinolytic. P-III class SVMPs reduced clotting time of human plasma. Fraction IIc was characterized as prothrombin activator and fraction Ic as factor X activator. In the absence of Ca2+, a firm clot was observed in chicken blood samples with fractions Ic, IIb and partially with fraction IIc. In contrast, without Ca2+, only fraction IIc was able to induce a firm clot in rat blood. In conclusion, functionally distinct forms of SVMPs were found in B. neuwiedi venom that affect distinct mechanisms in the coagulation system of humans, birds and small rodents. Distinct SVMPs appear to be more specialized to rat or chicken blood, strengthening the current hypothesis that toxin diversity enhances the possibilities of the snakes for hunting different prey or evading different predators. This functional diversity also impacts the complexity of human envenoming since different hemostatic mechanisms will be targeted by SVMPs accounting for the complexity of the response of humans to venoms.
    PLoS ONE 01/2014; 9(10):e109651. · 3.53 Impact Factor
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    ABSTRACT: Envenomation by Bothrops species results, among other symptoms, in haemostatic disturbances. These changes can be ascribed to the presence of enzymes, primarily serine proteinases some of which are structurally similar to thrombin and specifically cleave fibrinogen releasing fibrinopeptides. A rapid, three-step, chromatographic procedure was developed to routinely purify serine proteinases from the venoms of Bothrops alternatus and Bothrops moojeni. The serine proteinase from B. alternatus displays an apparent molecular mass of ∼32 kDa whereas the two closely related serine proteinases from B. moojeni display apparent molecular masses of ∼32 kDa and ∼35 kDa in SDS-PAGE gels. The partial sequences indicated that these enzymes share high identity with serine proteinases from the venoms of other Bothrops species. These proteins coagulate plasma and possess fibrinogenolytic activity but lack fibrinolytic activity.
    Toxicon 10/2013; · 2.92 Impact Factor
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    ABSTRACT: Snake venom proteomes/peptidomes are highly complex and subject to ontogenetic changes. Individual variation in the venom proteome of juvenile snakes is poorly known. We report the proteomic analysis of venoms from 21 juvenile specimens of Bothrops jararaca of different geographical origins and correlate it with the evaluation of important venom features. Individual venoms showed similar caseinolytic activities, however their amidolytic activities were significantly different. Rather intriguingly, plasma coagulant activity showed remarkable variability among the venoms but not the prothrombin-activating activity. LC-MS analysis showed significant differences between venoms, however, an interesting finding was the ubiquitous presence of the tripeptide ZKW, an endogenous inhibitor of metalloproteinases. Electrophoretic profiles of proteins submitted to reduction showed significant variability in total proteins, glycoproteins, and in the subproteomes of proteinases. Moreover, identification of differential bands revealed variation in most B. jararaca toxin classes. Profiles of venoms analyzed under non-reducing conditions showed less individual variability and identification of proteins in a conserved band revealed the presence of metalloproteinases and L-amino acid oxidase as common components of these venoms. Taken together, our findings suggest that individual venom proteome variability in B. jararaca exist since very early animal age and is not a result of ontogenetic and diet changes.
    Journal of Proteome Research 09/2013; · 5.06 Impact Factor
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    ABSTRACT: Snake venom serine proteinases (SVSPs) may affect hemostatic pathways by specifically activating components involved in coagulation, fibrinolysis and platelet aggregation or by unspecific proteolytic degradation. In this study, we purified and characterized a SVSP from Bothrops cotiara venom, named cotiarinase, which generated thrombin upon incubation with prothrombin. Cotiarinase was isolated by a two-step procedure including gel-filtration and cation-exchange chromatographies and showed a single protein band with a molecular mass of 29 kDa by SDS-polyacrylamide gel electrophoresis under reducing conditions. Identification of cotiarinase by mass spectrometric analysis revealed peptides that matched sequences of viperid SVSPs. Cotiarinase did not show fibrinogen-clotting, platelet-aggregating, fibrinogenolytic and factor X activating activities. Upon incubation with prothrombin the generation of thrombin was detected using the peptide substrate D-Phe-Pip-Arg-pNA. Moreover, mass spectrometric identification of prothrombin fragments generated by cotiarinase in the absence of co-factors (phospholipids, factor Va, factor Xa and Ca(2+) ions), indicated the limited proteolysis of this protein to release prothrombin 1, fragment 1 and thrombin. Cotiarinase is a novel SVSP that acts on prothrombin to release active thrombin that does not match any group of the current classification of snake venom prothrombin activators.
    Biochimie 04/2013; · 3.14 Impact Factor

Publication Stats

1k Citations
232.54 Total Impact Points

Institutions

  • 2003–2014
    • Instituto Butantan
      • • Laboratório de Fisiopatologia
      • • Laboratório Especial de Toxinologia Aplicada (LETA)
      • • Laboratório de Bioquímica e Biofísica
      San Paulo, São Paulo, Brazil
    • Showa University
      • Division of Pharmaceutical Sciences
      Shinagawa, Tōkyō, Japan
  • 2011
    • University of São Paulo
      • Departamento de Biologia Celular e do Desenvolvimento (ICB)
      Ribeirão Preto, Estado de Sao Paulo, Brazil
    • Instituto Adolfo Lutz
      • Divisão de Biologia Médica
      San Paulo, São Paulo, Brazil
  • 2003–2009
    • University of Virginia
      • Microbiology, Immunology and Cancer Biology (MIC)
      Charlottesville, VA, United States
  • 2006
    • Instituto Vital Brazil
      SRA, Rio Grande do Sul, Brazil
  • 2004
    • Institut Pasteur
      Lutetia Parisorum, Île-de-France, France