Characterization of the proinflammatory activity of a new serine protease (cdtsp-2) purified from the total venom of Crotalus durissus terrificus

• Grant number: 17/20291-0
• Support Opportunities: Regular Research Grants
• Start date: May 01, 2018
• End date: January 31, 2021
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Marcos Hikari Toyama
• Grantee: Marcos Hikari Toyama
• Host Institution: Instituto de Biociências (IB-CLP). Universidade Estadual Paulista (UNESP). Campus Experimental do Litoral Paulista. São Vicente , SP, Brazil
• Associated researchers: Marcos Antonio de Oliveira

Abstract

Recently our group isolated a new serine protease from the total venom of Crotalus durissus terrificus (cdtsp-2) as a molecular mass of 27 kDa, and its N-terminal sequence showed structural differences when compared to the gyroxin of Crotalus durissus terrificus, besides not presenting glycosylation such as gyroxin. Venom serine proteases (SVSP) interfere in the regulation and control of important biological reactions of blood homeostasis and can be classified as activators of the fibrinolytic system, procoagulant, anticoagulant and platelet aggregating enzymes. However, our preliminary results show that cdtsp-2 also presents significant edematogenic and myotoxic activity, which can be considered an unpublished fact and may introduce SVSPs as important agents capable of inducing pharmacological events at the cellular level as with sPLA2. In this project, we intend to characterize the pro-inflammatory mechanism and myotoxic activity induced by cdtsp-2 using the paw edema model in mice as the study model and using a combination of inhibitors and agonists related to the protease receptor signaling pathway (PARs ). On the other hand, two events can occur concomitantly, one related to the increase of reactive oxygen / nitrogen species that initiate an intracellular signaling cascade that may increase the expression of protein inflammatory mediators and the other is related to the activation of the acid cascade arachidonic. Thus, we also intend to establish a relationship between the activation of the PAR (s) receptors with the activation of the arachidonic acid cascade, monitoring the activity of cytosolic Fosfolipase A2, COX-2, Prostaglandin E2, TNFalpha, IL-6, IL- NFkb p65 and the relative concentration of Malonaldehyde (MDA).In addition to investigating the mechanism of CDT-2-induced mydynamic action of the venom of Crotalus durissus terrificus, this project will establish comparative bases using in silico analysis of cdtsp-2 of Crotalus durissus terrificus compared to gyroxin and thrombin, identifying structural differences that may be considered important for the differences between cdtsp-2 and gyroxin. The results obtained in this project can expand the action of the SVSPs to an agent of fundamental importance to the side of sPLA2 as agents inducing cellular and systemic events such as inflammation and myotoxicity and not only associated with coagulative changes. These results can help in the development of new therapeutic approaches and consider the contribution of antioxidant elements as enzymes, reassess some pain and inflammation drugs, focusing mainly on their antioxidant activity and the contribution of plant extracts. (AU)

Scientific publications

(15)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE MOURA, WERNER ALFINITO FEIO; SCHULTZ, LEONARDO; BREYER, CARLOS ALEXANDRE; DE OLIVEIRA, ANA LAURA PIRES; TAIRUM, CARLOS ABRUNHOSA; FERNANDES, GABRIELLA COSTA; TOYAMA, MARCOS HIKARI; PESSOA-JR, ADALBERTO; MONTEIRO, GISELE; DE OLIVEIRA, MARCOS ANTONIO. Functional and structural evaluation of the antileukaemic enzymel-asparaginase II expressed at low temperature by differentEscherichia colistrains. Biotechnology Letters, JUL 2020. (13/08617-7, 17/19942-7, 17/25272-4, 19/04054-4, 14/22039-9, 16/19245-1, 17/20291-0, 11/13500-6)

MANZI, AGATHA; DE-CARLI, BRUNO PAES; ROGGERO, AIRAM; DE MORAES, LAILA LUCYANE FERREIRA; ANNUNCIATO, ISABELLY; BELCHOR, MARIANA NOVO; NETO, DANIEL FERREIRA DE LIMA; DE OLIVEIRA, MARCOS ANTONIO; TOYAMA, MARCOS HIKARI. Theoretical evaluation of the malathion and its chemical derivatives interaction with cytosolic phospholipase A2 from zebrafish. Chemosphere, v. 311, p. 7-pg., 2023-01-01. (21/03352-1, 17/20291-0)

TOYAMA, MARCOS HIKARI; ROGERO, AIRAM; DE MORAES, LAILA LUCYANE FERREIRA; FERNANDES, GUSTAVO ANTONIO; DA CRUZ COSTA, CAROLINE RAMOS; BELCHOR, MARIANA NOVO; DE CARLI, AGATHA MANZI; DE OLIVEIRA, MARCOS ANTONIO. allic Acid as a Non-Selective Inhibitor of alpha/beta-Hydrolase Fold Enzymes Involved in the Inflammatory Process: The Two Sides of the Same Coi. HARMACEUTIC, v. 14, n. 2, FEB 2022. (20/03297-8, 17/19942-7, 17/20291-0)

DOS SANTOS, MELINA CARDOSO; TAIRUM, CARLOS ABRUNHOSA; MONTANHERO CABRERA, VITORIA ISABELA; GUIMARAES CAUZ, ANA CAROLINA; RIBEIRO, LUIZ FERNANDO; TOLEDO JUNIOR, JOSE CARLOS; TOYAMA, MARCOS HIKARI; GHILARDI LAGO, JOAO HENRIQUE; BROCCHI, MARCELO; SOARES NETTO, LUIS EDUARDO; et al. Adenanthin Is an Efficient Inhibitor of Peroxiredoxins from Pathogens, Inhibits Bacterial Growth, and Potentiates Antibiotic Activities. Chemical Research in Toxicology, v. N/A, p. 12-pg., 2022-05-10. (17/06263-4, 17/19942-7, 18/12628-8, 17/20291-0, 21/00209-3, 20/02868-1, 13/07937-8)

DOS SANTOS JUNIOR, ADEILSO BISPO; TAMAYOSE, CINTHIA I.; FERREIRA, MARCELO J. P.; BELCHOR, MARIANA N.; COSTA, CAROLINE R. C.; DE OLIVEIRA, MARCOS ANTONIO; TOYAMA, MARCOS HIKARI. Bioaffinity Fishing Procedure Using Secretory Phospholipase A2 for Screening for Bioactive Components: Modulation of Pharmacological Effect Induced by sPLA2 from Crotalus durissus terrificus by Hispidulin from Moquiniastrum floribundum. Molecules, v. 25, n. 2, JAN 2 2020. (17/20291-0, 14/21593-2, 17/19942-7, 14/20932-8)

BITTENCOURT RODRIGUES, CAROLINE FABRI; PENA FERREIRA, MARCELO JOSE; BELCHOR, MARIANA NOVO; COSTA, CAROLINE R. C.; NOVAES, DANIELLE P.; DOS SANTOS JUNIOR, ADEILSO BISPO; TAMAYOSE, CINTHIA I.; TERASHIMA PINHO, MARCUS VINICIUS; DE OLIVEIRA, MARCOS ANTONIO; TOYAMA, MARCOS HIKARI. Evaluation of the Inhibitory Potential of Casuarictin, an Ellagitannin Isolated from White Mangrove (Laguncularia racemosa) Leaves, on Snake Venom Secretory Phospholipase A2. MARINE DRUGS, v. 17, n. 7, JUL 2019. (17/20291-0, 14/20932-8, 14/21593-2, 13/10938-6)

ANNUNCIATO, I.; DE-CARLI, B. P.; MANZI, A.; NOVO BELCHOR, M.; DA CRUZ COSTA, C. R.; DE MORAES, L. L. F.; SANTOS SILVA, A. R. D.; DE OLIVEIRA, I. N.; DE OLIVEIRA, M. A.; TOYAMA, M. H.. PAF level regulation are crucial for embryonic development of zebrafish. FEBS OPEN BIO, v. 12, p. 1-pg., 2022-07-01. (17/20291-0)

COSTA, CAROLINE R. C.; BELCHOR, MARIANA NOVO; RODRIGUES, CAROLINE F. B.; TOYAMA, DANIELA DE OLIVEIRA; DE OLIVEIRA, MARCOS A.; NOVAES, DANIELLE P.; TOYAMA, MARCOS HIKARI. Edema Induced by a Crotalus durissus terrificus Venom Serine Protease (Cdtsp 2) Involves the PAR Pathway and PKC and PLC Activation. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 19, n. 8, p. 17-pg., 2018-08-01. (17/19942-7, 17/20291-0)

COSTA, CAROLINE R. C.; BELCHOR, MARIANA NOVO; RODRIGUES, CAROLINE F. B.; TOYAMA, DANIELA DE OLIVEIRA; DE OLIVEIRA, MARCOS A.; NOVAES, DANIELLE P.; TOYAMA, MARCOS HIKARI. Edema Induced by a Crotalus durissus terrificus Venom Serine Protease (Cdtsp 2) Involves the PAR Pathway and PKC and PLC Activation. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 19, n. 8, AUG 2018. (17/20291-0, 17/19942-7)

TAIRUM, CARLOS A.; SANTOS, MELINA CARDOSO; BREYER, CARLOS ALEXANDRE; PIRES DE OLIVEIRA, ANA LAURA; MONTANHERO CABRERA, VITORIA ISABELA; TOLEDO-SILVA, GUILHERME; MORI, GUSTAVO MARUYAMA; TOYAMA, MARCOS HIKARI; SOARES NETTO, LUIS EDUARDO; DE OLIVEIRA, MARCOS ANTONIO. Effects of Serine or Threonine in the Active Site of Typical 2-Cys Prx on Hyperoxidation Susceptibility and on Chaperone Activity. ANTIOXIDANTS, v. 10, n. 7, JUL 2021. (20/02868-1, 19/04054-4, 17/20291-0, 17/19942-7, 18/12628-8, 11/13500-6, 17/06263-4, 13/07937-8)

DE PAULA, CARLA PERES; DOS SANTOS, MELINA CARDOSO; TAIRUM, CARLOS A.; BREYER, CARLOS ALEXANDRE; TOLEDO-SILVA, GUILHERME; TOYAMA, MARCOS HIKARI; MORI, GUSTAVO MARUYAMA; DE OLIVEIRA, MARCOS ANTONIO. Glutaredoxin-like protein (GLP)-a novel bacteria sulfurtransferase that protects cells against cyanide and oxidative stresses. Applied Microbiology and Biotechnology, v. 104, n. 12, APR 2020. (13/16192-6, 13/07937-8, 17/19942-7, 17/06263-4, 11/13500-6, 17/20291-0, 10/00172-8, 10/16827-3)

GUILHEN LONGO, LARISSA VALLE; BREYER, CARLOS ALEXANDRE; NOVAES, GABRIELA MACHADO; GEGEMBAUER, GREGORY; LEITAO JR, NATANAEL PINHEIRO; OCTAVIANO, CARLA ELIZABETE; TOYAMA, MARCOS HIKARI; DE OLIVEIRA, MARCOS ANTONIO; PUCCIA, ROSANA. The Human PathogenParacoccidioides brasiliensisHas a Unique 1-Cys Peroxiredoxin That Localizes Both Intracellularly and at the Cell Surface. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 10, AUG 4 2020. (13/07937-8, 17/19942-7, 06/05095-6, 13/25950-1, 17/20291-0, 11/13500-6)

COSTA, CAROLINE R. C.; BELCHOR, MARIANA N.; ROGGERO, AIRAM; MORAES, LAILA L. L.; SAMELO, RICARDO; ANNUNCIATO, ISABELLY; BONTURI, CAMILA R. R.; OLIVA, MARIA L. V.; SOUSA, SERGIO F. F.; DE OLIVEIRA, MARCOS A. A.; et al. The First Anti-Snakebite and Hepatoprotective Characterization of a Trypsin Kunitz-like Inhibitor (EcTI) from the Plant Enterolobium contortisiliquum; A Case of Two Soul Mates Meeting. PHARMACEUTICALS, v. 16, n. 4, p. 17-pg., 2023-04-01. (03/10516-2, 17/19942-7, 17/20291-0)

CABRERA, VITORIA ISABELA MONTANHERO; SIVIDANES, GABRIELLE DO NASCIMENTO; QUINTILIANO, NATALIA FERNANDA; TOYAMA, MARCOS HIKARI; LAGO, JOAO HENRIQUE GHILARDI; DE OLIVEIRA, MARCOS ANTONIO. Exploring functional and structural features of chemically related natural prenylated hydroquinone and benzoic acid from Piper crassinervium (Piperaceae) on bacterial peroxiredoxin inhibition. PLoS One, v. 18, n. 2, p. 21-pg., 2023-02-24. (20/02868-1, 13/07937-8, 17/19942-7, 21/02789-7, 17/20291-0)

DE MOURA, WERNER ALFINITO FEIO; SCHULTZ, LEONARDO; BREYER, CARLOS ALEXANDRE; DE OLIVEIRA, ANA LAURA PIRES; TAIRUM, CARLOS ABRUNHOSA; FERNANDES, GABRIELLA COSTA; TOYAMA, MARCOS HIKARI; PESSOA-JR, ADALBERTO; MONTEIRO, GISELE; DE OLIVEIRA, MARCOS ANTONIO. Functional and structural evaluation of the antileukaemic enzymel-asparaginase II expressed at low temperature by differentEscherichia colistrains. Biotechnology Letters, v. 42, n. 11, p. 12-pg., 2020-07-07. (13/08617-7, 11/13500-6, 16/19245-1, 17/25272-4, 19/04054-4, 17/19942-7, 14/22039-9, 17/20291-0)