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Journal of Venomous Animals and Toxins including Tropical Diseases

On-line version ISSN 1678-9199

J. Venom. Anim. Toxins incl. Trop. Dis vol.13 no.4 Botucatu  2007

http://dx.doi.org/10.1590/S1678-91992007000400003 

REVIEW ARTICLE

 

Professor José Roberto Giglio and toxinnology in Brazil: 48 years in research

 

 

Soares A. M.I; Oliveira C. Z.I; Santana C. D.I; Menaldo D. L.I; Silveira L. B.I; Teixeira S. S.I; Rueda A. Q.I; Marcussi S.I, II

IDepartment of Clinical, Toxicological and Bromatological Analyses, School of Pharmaceutical Sciences-FCFRP, University of São Paulo-USP, Ribeirão Preto, São Paulo State, Brazil
II
Department of Biochemistry and Immunology, Ribeirão Preto School of Medicine-FMRP, USP, Ribeirão Preto, São Paulo State, Brazil

Correspondence to

 

 


ABSTRACT

This work succinctly describes the professional and scientific life of Dr. José R. Giglio, one of the most outstanding Brazilian researchers in the field of Toxinology. During his long and successful career, he has made major contributions, especially in elucidating the function, structure, and mechanisms of action of animal venom proteins (from snakes, scorpions and spiders) as well as the characterization of antibodies and several inhibitors of venoms and toxins. We present here a brief history of Dr. Giglio’s personal and professional life, also reporting some of his numerous published scientific articles on venoms from snakes (Bothrops, Crotalus, and other genera), scorpions (Tityus sp), spiders (Phoneutria sp), their isolated toxins and natural inhibitors. Thus, this work is a tribute to Dr. Giglio in his 73rd birthday, having devoted 48 years of his life studying animal venoms, an effort that has continued even after his formal retirement from university duties.

Key words: Toxinology, animal venoms, toxins, inhibitors, homage, Giglio JR.


 

 

INTRODUCTION

Dr. José Roberto Giglio (Figure 1) is one of the most outstanding Brazilian toxinologists. During his long and successful career, he has contributed a lot, especially with the elucidation of the function, structure, and mechanism of action of proteins from snake and scorpion venoms, and with the characterization of antibodies and several inhibitors of these venoms and toxins. He is an educator who was directly involved in the creation of the Department of Biochemistry and Immunology at the University of São Paulo (FMRP-USP), Brazil, and was responsible for the formation of several researchers who have made relevant contributions to scientific research. He is an outstanding citizen of the academic community, and despite his tight schedule, he never refused to take on another professional obligation that would promote the development of science. Through his loyalty and generosity, he has become a true friend to many of us.

 

 

Giglio was born in 1934 at the city of São Paulo, Brazil. In 1957, he graduated in Chemistry at the School of Philosophy, Languages and Humana Sciences, University of São Paulo [Faculdade de Filosofia Ciências e Letras, Universidade de São Paulo-USP] among a selected group of only five students, out of 30 that were enrolled in 1954. He was professor of Chemistry at the Oswaldo Cruz School [Colégio Oswaldo Cruz] (1957-58), and worked in the Laboratory of Chemistry at São Paulo Cancer Hospital [Hospital do Câncer de São Paulo]. In 1959, Giglio started to work at the Biochemistry Department of the Ribeirão Preto School of Medicine [Faculdade de Medicina de Ribeirão Preto, (FMRP-USP)] with Dr. José Moura Gonçalves as Professor of Biochemistry where he began his first studies on snake venom and where he met his wife Ms. Albertina Exposto Giglio. They married in January 1962 and lived in the USP Campus for 40 years. Dr. Giglio created a solid family with sons and grandsons simultaneously with the development of his academic career.

Starting his PhD program in 1959 with the project entitled "Terminal amino acids of crotamine". Giglio obtained his Doctor degree in Biochemistry in 1962 at USP under the guidance of Dr. José M. Gonçalves. In his first time abroad, he learned to analyze amino acids, becoming himself the responsible for the purification and determination of crotamine amino acids composition, being the first one to accomplish this kind of analysis in Brazil.

Dr. Giglio started his continuous and productive scientific contribution, publishing in the Anais da Academia Brasileira de Letras, with his supervisor Dr. José M. Gonçalves, his first complete article about spectrophotometric studies of the interaction between bovine serum albumin and pentachlorophenol (27). In 1965, Giglio had a post-doctoral training at Cornell University, USA, working on the chemistry of biomacromolecules, with studies dealing with the primary structure of thrombin.

In 1969, he published his first article in an international journal on the activation of bovine prothrombin by trypsin and on the identification of the residues from the N-terminal sequence of that protein, a work developed with his first doctoral student, Antonio Rossi (currently Professor at the Medical School FMRP-USP). Dr. Giglio published some articles related to bovine and porcine prothrombin and thrombin (26, 28). From 1975 to 1976, he worked at the Imperial College of London as Visiting Professor, where he got familiar with the manual sequencing of proteins, employing ribitol dehydrogenase, an important biotechnology protein obtained from Klebsiella aerogenes (22). From his results, Dr. Giglio and colleagues published the complete sequence of a mutant enzyme showing a higher xylitol dehydrogenase activity (31).

The first studies performed by Dr. Giglio on proteins from snake venoms were published in 1975 and dealt with the analytical investigation of crotamine, a basic neurotoxic peptide (Mr~ 4,880) isolated from South American rattlesnake Crotalus d. terrificus venom (25), and in 1982, with studies of RAMAN spectra from the same protein which allowed him to conclude that the only Tyr residue was buried, and the Trp residues were exposed to the solvent.

Giglio became affiliated with USP as a Full Professor in 1990, devoting his life to teaching, learning and performing research, preparing post-graduate and graduate students, forming PhDs, and building a community of researchers and disciples. There are several renowned researchers who were guided by Dr. Giglio, such as Dr. Suely V. Sampaio (current Rector of USP and the first woman to occupy this position), Eliane C. Arantes, Andreimar M. Soares, Suzelei C. França, Veridiana M. Rodrigues, Maria Ines Homsi-Brandeburgo, Alessandra L. Cecchini, Silvia H. Andrião-Escarso, Adélia C. O. Cintra, Heloísa S. Selistre-de-Araújo, Antônio Rossi and others (PhDs, Full Professors of Federal or State Universities). His guidance was very important for their personal and professional maturing process. Since 1983, Dr. Giglio has been supported in his research by two excellent technicians and friends, Carlos A. Vieira and Odete A. B. Cunha, who have given him technical support of research and have played a relevant role in his several scientific publications.

Since the end of the 80’s until now, Dr. Giglio has performed several studies on snake venom proteins involving their isolation and their biochemical, structural and functional characterization. Searching of new methods to study proteins, which developed concomitantly with the admission of graduate students at Dr. Giglio's Laboratory, and the contribution of other researchers from different areas created a fruitful and stimulating atmosphere for the study of these proteins following new tendencies based on the evolution in Biochemistry and resulting in the elucidation of their structure-function relationships.

 

SNAKE VENOMS: TOXINS AND INHIBITORS

The first protein to have its complete amino acid sequence accomplished in Brazil was BthTX-I (Figure 2A), a basic PLA2 homologue with myotoxic activity and Mr~13,720, isolated from Bothrops jararacussu venom (20, 30); in addition, the antagonism of this myotoxic protein by polyanions was investigated (49). The possible role of the enzymatic activity of BthTX-I on neuromuscular effects induced by this myotoxin, as well as its crystallization and preliminary X-ray diffraction, was investigated in 1995 (Figure 2B). Conformational changes in BthTX-I were observed by spectroscopy studies (70), whereas mast cell degranulation induced by BthTXs native and chemically modified with p-bromophenacyl bromide (BPB) demonstrated the dissociation between enzymatic and pharmacological activities (37). Oliveira et al. (52) accomplished an important study with BthTX-I and discovered that the dimer, a component of protein structure, is essential for its capability to cause membrane lesions independently of Ca2+. In this same year, the position of the disulfide bridges of BthTX-I structure was determined (Figure 2A).

 


 

The functional and structural characterization of an acidic PLA2 (Mr~13,700) composed of 122 amino acid residues exerting hypotensive and inhibitory activities of platelet aggregation, isolated from B. jararacussu venom, was performed (2), as well as the elucidation of the sequence of nerve growth factor (Bj-NGF) by Molecular Biology (35). In 2003, isolation and characterization of biological activities of other acidic PLA2 isoforms from B. jararacussu venom were accomplished (36). In 2004, the analysis of venom gland transcriptome of B. jararacussu paved the way for studies on the elucidation of the relationships between PLA2s function and structure (34). Other studies included the evaluation of presynaptic activity of BthTX-I (55); the cloning and identification of cDNA of acidic phospholipase A2 with bactericidal and antitumoral effects from B. jararacussu (60, 61); and the study of signal transduction via platelet aggregation induced by BthTX-II, applied in its native and modified form (23). This BthTX-II enzyme, a basic PLA2 Asp49 previously isolated from the same venom (30), showing low catalytic activity, was crystallized and its primary structure completely elucidated (57).

Several other studies were developed using these isolated proteins from Bothrops and Crotalus snake venoms (3, 9, 11, 17, 18, 24, 38-43, 50, 53, 64, 73-75, 77, 78, 80, 83, 87, 88).

Despite an emphasis on the study of PLA2, other protein types were also investigated by Dr. Giglio and colleagues, such as the isolation and characterization of a new coagulant factor from B. jararacussu venom which is a glycoprotein with fibrinogenolytic activity (1). Recently, BjussuSP-I, another thrombin-like serine protease isolated from B. jararacussu venom with Mr~61,000 and pI~3.8 was also isolated and functionally characterized (69). Metalloproteases are other types of proteins that have been studied by him. Neuwiedase, a non-hemorrhagic metalloprotease isolated from B. neuwiedi with fibrinogenolytic activity (Mr~20,000 and pI 5.9) was characterized (62, 63). Recently, BjussuMP-II, a non-hemorrhagic metalloprotease P-I (Mr~24,000 and pI 6.0) was isolated from B. jararacussu venom (47).

Dr. Giglio also coauthored four review articles, being two of them on phospholipases A2, one on PLA2 inhibitors and another one on medicinal plants with anti-snake venom properties. The first one described the effect of chemical modifications of different amino acid residues on PLA2s of snake venoms, especially Bothrops, and their effects on the enzymatic, pharmacological and toxic activities induced by these enzymes (76). The second review covered myotoxic phospholipases A2 from Bothrops snake venom describing the structure-function of these enzymes, and paving the way for future perspectives in the study of these enzymes (81).

Due to the great number of venom proteins isolated and characterized, the search for new methods and compounds for an improved therapy of snakebite envenomation has been undertaken by Dr. Giglio in his studies (15, 16, 32, 54, 56, 71, 72, 79, 80, 86, 89, 92). Dr. Giglio also participated in other two review articles describing the potential of medicinal plants against snake venoms and isolated toxins (82) and on inhibitors of snake venom PLA2s (48).

More recently, he and others described the utilization of a library of human recombinant antibodies called Griffin.1 for fragment selection capable of inhibiting the myotoxic activity of PLA2 from B. jararacussu venom (84). Now, another library is being described for recombinant antibodies capable of recognizing and inhibiting the pharmacological and enzymatic activities induced by Crotalus d. terrificus CB (53).

 

OTHER ANIMAL VENOMS

Another subject explored by Dr. Giglio, his students and collaborators, was the study of scorpion venom toxins, mainly from species of the genus Tityus (5-8, 10, 14, 19, 21, 29, 33, 39, 45, 46, 51, 58, 65-68, 85, 90, 91).

Fractions of Phoneutria nigriventer spider venom were studied by Dr. Giglio and his staff, as well as the activation of kallikrein-kininogen-kinin system in rabbit skin and the in vivo increase of microvascular permeability through this system (4, 13). A polypeptide from P. nigriventer venom was studied and showed a short-duration dose-dependent stimulation in the contraction of arterial and rabbit venous vessels (12). Marangoni et al. (44) described PNV1 as a protein from P. nigriventer venom with spasmogenic activity in the smooth muscle of rabbit vessels. Also, a polypeptide that acts relaxing cavernous bodies of rabbit in vitro was isolated and partially characterized (59).

 

CONCLUSION

In his career as professor, researcher and guide, Dr. Giglio published more than 130 articles in international journals of Biochemistry and Pharmacology and still has many works in development and/or submitted to scientific journals. He collaborated in projects developed by important research groups at USP and scientists of other national and international institutions. On May 21, 1995, Folha de São Paulo, one of the most distinguished Brazilian newspapers, published the names of 170 Brazilian scientists whose works caused great impact in the scientific world from 1981 to 1993 (ISI database, USA); Dr. Giglio was included in this selected group. He also had the recognition of CNPq (National Council for Scientific and Technological Development), being honored as Emeritus Researcher. He has plenty of qualities, such as dignity, courage, sensibility and love to his profession, qualities that he maintained along his entire life, even after his formal retirement. After such a long and successful career, he should be full of satisfaction for the duty performed. However, as all great scientists, he continues performing research and helping those who look for his advice and support.

 

ACKNOWLEDGMENTS

The authors are grateful to Dr. J. M. Gutiérrez (ICP, Costa Rica) for critical reading of this manuscript and, CNPq, CAPES and FAPESP for financial support.

 

REFERENCES

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2 ANDRIAO-ESCARSO SH., SOARES AM., FONTES MR., FULY AL., CORREA FM., ROSA JC., GREENE LJ., GIGLIO JR. Structural and functional characterization of an acidic platelet aggregation inhibitor and hypotensive phospholipase A2 from Bothrops jararacussu snake venom. Biochem. Pharmacol., 2002, 64, 723-32.         [ Links ]

3 ANDRIAO-ESCARSO SH., SOARES AM., RODRIGUES VM., ANGULO Y., DIAZ C., LOMONTE B., GUTIERREZ JM., GIGLIO JR. Myotoxic phospholipases A2 in Bothrops snake venoms: effect of chemical modifications on the enzymatic and pharmacological properties of bothropstoxins from Bothrops jararacussu. Biochimie, 2000, 82, 755-63.         [ Links ]

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5 ARANTES EC., PRADO WA., SAMPAIO SV., GIGLIO JR. A simplified procedure for the fractionation of Tityus serrulatus venom: isolation and partial characterization of TsTX-IV, a new neurotoxin. Toxicon, 1989, 27, 907-16.         [ Links ]

6 ARANTES EC., RICCIOPPO NETO F., SAMPAIO SV., VIEIRA CA., GIGLIO JR. Isolation and characterization of TsTX-V, a new neurotoxin from Tityus serrulatus scorpion venom which delays the inactivation of Na+ channels. Biochim. Biophys. Acta, 1994, 1199, 69-75.        [ Links ]

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8 AREAS EP., GIGLIO JR., ARANTES EC., KAWANO Y. Raman and infrared spectra of toxin gamma from the venom of the scorpion Tityus serrulatus. Biochim. Biophys. Acta, 1987, 915, 292-8.         [ Links ]

9 AZEVEDO JR WF., WARD RJ., CANDURI F., SOARES A., GIGLIO JR., ARNI RK. Crystal structure of piratoxin-I: a calcium-independent, myotoxic phospholipase A2- homologue from Bothrops pirajai venom. Toxicon, 1998, 36, 1395-406.         [ Links ]

10 BARHANIN J., ILDEFONSE M., ROUGIER O., SAMPAIO SV., GIGLIO JR. Tityus gamma toxin, a high-affinity effector of the Na+ channel in muscle, with a selectivity for channels in the surface membrane. Pflugers Arch., 1984, 400, 22-7.         [ Links ]

11 BATINA MF., CINTRA AC., VERONESE EL., LAVRADOR MA., GIGLIO JR., PEREIRA PS., DIAS DA., FRANCA SC., SAMPAIO SV. Inhibition of the lethal and myotoxic activities of Crotalus durissus terrificus venom by Tabernaemontana catharinensis: identification of one of the active components. Planta Med., 2000, 66, 424-8.         [ Links ]

12 BENTO AC., NOVELLO JC., MARANGONI S., ANTUNES E., GIGLIO JR., OLIVEIRA B., NUCCI G. Identification of a new vascular smooth muscle contracting polypeptide in Phoneutria nigriventer spider venom. Biochem Pharmacol., 1993, 46, 1092-5.         [ Links ]

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14 BORGES MH., ARANTES EC., GIGLIO JR. Isolation and characterization of toxic proteins from the venom of the Venezuelan scorpion Tityus discrepans (Karsch). Toxicon, 1990, 28, 1011-7.         [ Links ]

15 BORGES, MH., SOARES AM., RODRIGUES VM., ANDRIAO-ESCARSO SH., DINIZ H., HAMAGUCHI A., QUINTERO A., LIZANO S., GUTIERREZ JM., GIGLIO JR., HOMSI-BRANDEBURGO MI. Effects of aqueous extract of Casearia sylvestris (Flacourtiaceae) on actions of snake and bee venoms and on activity of phospholipases A2. Comp. Biochem. Physiol., 2000, 127B, 21-30.        [ Links ]

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17 CASTRO RC., LANDUCCI EC., TOYAMA MH., GIGLIO JR., MARANGONI S., NUCCI G., ANTUNES E. Leukocyte recruitment induced by type-II phospholipases A2 into the rat pleural cavity. Toxicon, 2000, 38, 1773-85.         [ Links ]

18 CECCHINI AL., SOARES AM., CECCHINI R., OLIVEIRA AH., WARD RJ., GIGLIO JR., ARANTES EC. Effect of crotapotin on the biological activity of Asp49 and Lys49 phospholipases A2 from Bothrops snake venoms. Comp. Biochem. Physiol., 2004, 138C, 429-36.         [ Links ]

19 CECCHINI AL., VASCONCELOS F., AMARA SG., GIGLIO JR., ARANTES EC. Effects of Tityus serrulatus scorpion venom and its toxin TsTX-V on neurotransmitter uptake in vitro. Toxicol. Appl. Pharmacol., 2006, 217, 196-203.         [ Links ]

20 CINTRA AC., MARANGONI S., OLIVEIRA B., GIGLIO JR. Bothropstoxin-I: amino acid sequence and function. J. Protein Chem., 1993, 12, 57-64.         [ Links ]

21 CORREA MM., SAMPAIO SV., LOPES RA., MANCUSO LC., CUNHA AO., FRANCO JJ., GIGLIO JR. Biochemical and histopathological alterations induced in rats by Tityus serrulatus scorpion venom and its major neurotoxin tityustoxin-I. Toxicon, 1997, 35, 1053-67.         [ Links ]

22 DOTHIE JM., GIGLIO JR., MOORE CB., TAYLOR SS., HARTLEY BS. Ribitol dehydrogenase of Klebsiella aerogenes. Biochem. J., 1985, 230, 569-78.         [ Links ]

23 FULY AL., SOARES AM., MARCUSSI S., GIGLIO JR., GUIMARAES JA. Signal transduction pathways involved in the platelet aggregation induced by a D-49 phospholipase A2 isolated from Bothrops jararacussu snake venom. Biochimie, 2004, 86, 731-9.         [ Links ]

24 GAMBERO A., LANDUCCI EC., TOYAMA MH., MARANGONI S., GIGLIO JR., NADER HB., DIETRICH CP., NUCCI G., ANTUNES E. Human neutrophil migration in vitro induced by secretory phospholipases A2: a role for cell surface glycosaminoglycans. Biochem. Pharmacol., 2002, 63, 65-72.         [ Links ]

25 GIGLIO JR. Analytical studies on crotamine hydrochloride. Anal. Biochem., 1975, 69, 207-21.         [ Links ]

26 GIGLIO JR. Identification of N-terminal tryptophan in peptides. Anal. Biochem. 1977, 82, 262-4.        [ Links ]

27 GIGLIO JR., GONÇALVES JM. Estudo espectrofotométrico da interação entre soro-albumina bovina e pentaclorofenol. An. Acad. Bras. Ci., 1963, 35, 293-8.         [ Links ]

28 GIGLIO JR., ROSSI A., LEONE FA., CHIERICATO G., SAY JC. Isolation and characterization of an active three-chain molecular species of bovine thrombin. Biochem. J., 1976, 159, 29-33.         [ Links ]

29 GONÇALVES AA., TOYAMA MH., CARNEIRO EM., MARANGONI S., ARANTES EC., GIGLIO JR., BOSCHERO AC. Participation of Na(+) channels in the potentiation by Tityus serrulatus alpha-toxin TsTx-V of glucose-induced electrical activity and insulin secretion in rodent islet beta-cells. Toxicon, 2003, 41, 1039-45.         [ Links ]

30 HOMSI-BRANDEBURGO MI., QUEIROZ LS., SANTO-NETO H., RODRIGUESSIMIONI L., GIGLIO JR. Fractionation of Bothrops jararacussu snake venom: partial chemical characterization and biological activity of bothropstoxin. Toxicon, 1988, 26, 615-27.         [ Links ]

31 HOMSI-BRANDEBURGO MI., TOYAMA MH., MARANGONI S., WARD RJ., GIGLIO JR., HARTLEY BS. The amino acid sequence of ribitol dehydrogenase-F, a mutant enzyme with improved xylitol dehydrogenase activity. J. Protein Chem., 1999, 18, 489-95.         [ Links ]

32 IZIDORO LF., RODRIGUES VM., RODRIGUES RS., FERRO EV., HAMAGUCHI A., GIGLIO JR., HOMSI-BRANDEBURGO MI. Neutralization of some hematological and hemostatic alterations induced by neuwiedase, a metalloproteinase isolated from Bothrops neuwiedi pauloensis snake venom, by the aqueous extract from Casearia mariquitensis (Flacourtiaceae). Biochimie, 2003, 85, 669-75.         [ Links ]

33 JONAS P., VOGEL W., ARANTES EC., GIGLIO JR. Toxin gamma of the scorpion Tityus serrulatus modifies both activation and inactivation of sodium permeability of nerve membrane. Pflugers Arch., 1986, 407, 92-9.         [ Links ]

34 KASHIMA S., ROBERTO PG., SOARES AM., ASTOLFI-FILHO S., PEREIRA JO., GIULIATI S., FARIA JR M., XAVIER MA., FONTES MR., GIGLIO JR., FRANCA SC. Analysis of Bothrops jararacussu venomous gland transcriptome focusing on structural and functional aspects: I-gene expression profile of highly expressed phospholipases A2. Biochimie, 2004, 86, 211-9.         [ Links ]

35 KASHIMA S., SOARES AM., ROBERTO PG., PEREIRA JO., ASTOLFI-FILHO S., CINTRA AO., FONTES MR., GIGLIO JR., FRANCA SC. cDNA sequence and molecular modeling of a nerve growth factor from Bothrops jararacussu venomous gland. Biochimie, 2002, 84, 675-80.        [ Links ]

36 KETELHUT DF., MELLO MH., VERONESE EL., ESMERALDINO LE., MURAKAMI MT., ARNI RK., GIGLIO JR., CINTRA AC., SAMPAIO SV. Isolation, characterization and biological activity of acidic phospholipase A2 isoforms from Bothrops jararacussu snake venom. Biochimie, 2003, 85, 983-91.         [ Links ]

37 LANDUCCI EC., CASTRO RC., PEREIRA MF., CINTRA AC., GIGLIO JR., MARANGONI S., OLIVEIRA B., CIRINO G., ANTUNES E., NUCCI G. Mast cell degranulation induced by two phospholipase A2 homologues: dissociation between enzymatic and biological activities. Eur. J. Pharmacol., 1998, 343, 257-63.         [ Links ]

38 LANDUCCI EC., CASTRO RC., TOYAMA M., GIGLIO JR., MARANGONI S., NUCCI G., ANTUNES E. Inflammatory oedema induced by the lys-49 phospholipase A2 homologue piratoxin-i in the rat and rabbit. Effect of polyanions and pbromophenacyl bromide. Biochem. Pharmacol., 2000, 59, 1289-94.         [ Links ]

39 LIBERONA JL., CAVIEDES P., TASCON S., HIDALGO J., GIGLIO JR., SAMPAIO SV., CAVIEDES R., JAIMOVICH E. Expression of ion channels during differentiation of a human skeletal muscle cell line. J. Muscle Res. Cell Motil., 1997, 18, 587-98.         [ Links ]

40 LOMONTE B., ANGULO Y., RUFINI S., CHO W., GIGLIO JR., OHNO M., DANIELE JJ., GEOGHEGAN P., GUTIERREZ JM. Comparative study of the cytolytic activity of myotoxic phospholipases A2 on mouse endothelial (tEnd) and skeletal muscle (C2C12) cells in vitro. Toxicon, 1999, 37, 145-58.         [ Links ]

41 MAGRO AJ., SOARES AM., GIGLIO JR., FONTES MR. Crystal structures of BnSP-7 and BnSP-6, two Lys49-phospholipases A2: quaternary structure and inhibition mechanism insights. Biochem. Biophys. Res. Commun., 2003, 311, 713-20.         [ Links ]

42 MANCIN AC., SOARES AM., ANDRIAO-ESCARSO SH., FACA VM., GREENE LJ., ZUCCOLOTTO S., PELA IR., GIGLIO JR. The analgesic activity of crotamine, a neurotoxin from Crotalus durissus terrificus (South American rattlesnake) venom: a biochemical and pharmacological study. Toxicon, 1998, 36, 1927-37.         [ Links ]

43 MANCUSO LC., CORREA MM., VIEIRA CA., CUNHA OA., LACHAT JJ., SELISTRE-DE-ARAÚJO HS., OWNBY CL., GIGLIO JR. Fractionation of Bothrops pirajai snake venom: isolation and characterization of piratoxin-I, a new myotoxic protein. Toxicon, 1995, 33, 615-26.        [ Links ]

44 MARANGONI S., BORGES NC., MARANGONI RA., ANTUNES E., VIEIRA CA., NOVELLO JC., DOMONT GB., GIGLIO JR., OLIVEIRA B., NUCCI G. Biochemical characterization of a vascular smooth muscle contracting polypeptide purified from Phoneutria nigriventer (armed spider) venom. Toxicon, 1993, 31, 377-84.         [ Links ]

45 MARANGONI S., GHISO J., SAMPAIO SV., ARANTES EC., GIGLIO JR., OLIVEIRA B., FRANGIONE B. The complete amino acid sequence of toxin TsTX-VI isolated from the venom of the scorpion Tityus serrulatus. J. Protein Chem., 1990, 9, 595-601.         [ Links ]

46 MARANGONI S., TOYAMA MH., ARANTES EC., GIGLIO JR., SILVA CA., CARNEIRO EM., GONÇALVES AA., OLIVEIRA B. Amino acid sequence of TsTX-V, an alpha-toxin from Tityus serrulatus scorpion venom, and its effect on K+ permeability of beta-cells from isolated rat islets of Langerhans. Biochim. Biophys. Acta., 1995, 1243, 309-14.         [ Links ]

47 MARCUSSI S., OLIVEIRA CZ., MAZZI MV., MENALDO DL., FULY AL., GOMES OA., MAGRO AJ., FONTES MM., GIGLIO JR., SOARES AM. A New nonhemorrhagic metalloprotease from Bothrops jararacussu snake venom: Functional and structural characterization. Biochim. Biophys. Acta, 2007.         [ Links ]

48 MARCUSSI S., SANT’ANA CD., OLIVEIRA CZ., RUEDA AQ., MENALDO DL., BELEBONI RO., STÁBELI RG., GIGLIO JR., FONTES MR., SOARES AM. Snake venom phospholipase A2 inhibitors: medicinal chemistry and therapeutic potential. Curr. Top. Med. Chem., 2007b, 7, 743-56.         [ Links ]

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50 NICASTRO G., FRANZONI L., CHIARA C., MANCIN AC., GIGLIO JR., SPISNI A. Solution structure of crotamine, a Na+ channel affecting toxin from Crotalus durissus terrificus venom. Eur. J. Biochem., 2003, 270, 1969-79.         [ Links ]

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Correspondence to:
Andreimar Martins Soares Faculdade de Ciências Farmacêuticas de Ribeirão Preto Av. do Café s/n, Campus Universitário da USP
14040-903, Ribeirão Preto, SP, Brasil
Phone: +55 16 3602-4714; Fax: +55 16 3602-4725
Email: andreims@fcfrp.usp.br

Received: August 10, 2007
Accepted: August 15, 2007
Abstract published online: August 27, 2007 Full paper published online: November 30, 2007
Conflicts of interest: There is no conflict.

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