[en] In this study we expressed the Ts8, a neurotoxin from Tityus serrulatus scorpion venom, in Pichia pastoris yeast. We evaluated the peptide expression in different conditions, such as pH, temperature, and addition of casamino acids supplement. Analyses of expressed products by mass spectrometry and Edman degradation showed that rTs8 has sites that allow its cleavage by yeast proteases released into the culture medium. The casamino acids addition was favourable for toxin expression, however, was not sufficient to minimize proteolytic degradation. Functional assays with recombinant toxin fragments and native toxins have demonstrated the release of cytokines such as TNF-α and IL-1β in some peptides tested. In addition, the toxins were shown to inhibit the Pichia pastoris growth in antifungal test and were not toxic to alveolar macrophages cells at the concentrations analyzed The electrophysiological screening, by voltage clamp technique, showed that the rTs8 fragment with the highest molecular weight inhibited the Kv1.3 channel, whereas the N-terminal fragment had no activity on the ion channels tested.
Disciplines :
Chemistry
Author, co-author :
Cordeiro, Francielle Almeida; Department of BioMolecular Sciences, Faculty of Pharmaceutical Sciences of Ribeirao Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil. Electronic address: fran_ac@alumni.usp.br
Gobbi Amorim, Fernanda ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Boldrini-França, Johara; School of Biochemistry, Biomedical Sciences, University of Bristol, 1 Tankard's Close, Bristol, BS8 1TD, UK
Pinheiro-Júnior, Ernesto Lopes; Toxicology and Pharmacology, KU Leuven, O&N II Herestraat 49 - PO Box 922, 3000, Leuven, Belgium
Cardoso, Iara Aimê; Department of BioMolecular Sciences, Faculty of Pharmaceutical Sciences of Ribeirao Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil
Zoccal, Karina Furlani; Center University Barão of Mauá, St. Ramos of Azavedo, N 423, 14090-062, Ribeirão Preto, SP, Brazil
Peigneur, Steve; Toxicology and Pharmacology, KU Leuven, O&N II Herestraat 49 - PO Box 922, 3000, Leuven, Belgium
Faccioli, Lucia Helena; Department of Clinical Analyses, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil
Tytgat, Jan; Toxicology and Pharmacology, KU Leuven, O&N II Herestraat 49 - PO Box 922, 3000, Leuven, Belgium
Arantes, Eliane Candiani; Department of BioMolecular Sciences, Faculty of Pharmaceutical Sciences of Ribeirao Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil. Electronic address: ecabraga@fcfrp.usp.br
Language :
English
Title :
Heterologous expression of Ts8, a neurotoxin from Tityus serrulatus venom, evidences its antifungal activity.
CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico [BR] FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo [BR] FWO - Fonds Wetenschappelijk Onderzoek Vlaanderen [BE] CAPES - Coordenação de Aperfeicoamento de Pessoal de Nível Superior [BR]
Funding text :
This study received financial support from Conselho Nacional de Desenvolvimento Científico e Tecnológico ( CNPq , The National Council for Scientific and Technological Development , scholarship to FAC - 155,276/2018–2 and grant to ECA - 306,479/2017–6 ), Fundacao de Amparo a Pesquisa do Estado de Sao Paulo ( FAPESP , Sao Paulo Research Foundation , grant to ECA - 2019/10,173–6 and scholarship to ELPJ - 2016/04761–4 ), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ( CAPES , Coordination for the Improvement of Higher Education Personnel , This research was also funded by FWO - Vlaanderen grants GOA4919N , GOE7120N and GOC2319N awarded to J.T. and 12W7822N to S.P.This study received financial support from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, The National Council for Scientific and Technological Development, scholarship to FAC - 155,276/2018–2 and grant to ECA - 306,479/2017–6), Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP, Sao Paulo Research Foundation, grant to ECA - 2019/10,173–6 and scholarship to ELPJ - 2016/04761–4), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Coordination for the Improvement of Higher Education Personnel, This research was also funded by FWO-Vlaanderen grants GOA4919N, GOE7120N and GOC2319N awarded to J.T. and 12W7822N to S.P.
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