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3. Molecular cloning, expression and IgE-immunoreactivity of phospholipase A1, a major allergen from Polybia paulista (Hymenoptera: Vespidae) venom

Author

Perez-Riverol, Amilcar, Campos Pereira, Franco Dani, Musacchio Lasa, Alexis, Romani Fernandes, Luis Gustavo, Santos-Pinto, José Roberto Aparecido dos, Justo-Jacomini, Débora Lais, Oliveira de Azevedo, Gabriel, Bazon, Murilo Luiz, Palma, Mario Sergio, Zollner, Ricardo de Lima, and Brochetto-Braga, Márcia Regina

Published
2016
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4. Gadolinium(Iii) Oxide Nanoparticles Conjugated with Amylovis® with Applications in the Early Detection of Alzheimer's Disease. Part Ii

Author

Guerrero-Porras, Gabriel Rafael, primary, Díaz-García, Alicia M., additional, González-Dalmau, Evelio R., additional, Hernández-Rodríguez, Armando J., additional, Reyes-Molina, Israel, additional, Musacchio-Lasa, Alexis, additional, Espinosa-Rodríguez, Luis A., additional, Rodriguez-Tanty, Chryslaine, additional, and Sablón-Carrazana, Marquiza, additional

Published
2022
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5. The Combination of the CIGB-300 Anticancer Peptide and Cisplatin Modulates Proteins Related to Cell Survival, DNA Repair and Metastasis in a Lung Cancer Cell Line Model

Author

Rodríguez-Ulloa, Arielis, primary, Ramos, Yassel, additional, Sánchez-Puente, Aniel, additional, Perera, Yasser, additional, Musacchio-Lasa, Alexis, additional, Fernández-de-Cossio, Jorge, additional, Padrón, Gabriel, additional, López, Luis J.G., additional, Besada, Vladimir, additional, and Perea, Silvio E., additional

Published
2019
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6. Phospholipase A1-based cross-reactivity among venoms of clinically relevant Hymenoptera from Neotropical and temperate regions

Author

Perez-Riverol, Amilcar, primary, Fernandes, Luís Gustavo Romani, additional, Musacchio Lasa, Alexis, additional, dos Santos-Pinto, José Roberto Aparecido, additional, Moitinho Abram, Débora, additional, Izuka Moraes, Gabriel Hideki, additional, Jabs, Frederic, additional, Miehe, Michaela, additional, Seismman, Henning, additional, Palma, Mario Sergio, additional, de Lima Zollner, Ricardo, additional, Spillner, Edzard, additional, and Brochetto-Braga, Márcia Regina, additional

Published
2018
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7. Indirect and competitive enzyme-linked immunosorbent assays for monitoring the humoral response against human VEGF

Author

Sánchez Ramírez, Javier, primary, Morera Díaz, Yanelys, additional, Musacchio Lasa, Alexis, additional, Bequet-Romero, Mónica, additional, Muñoz Pozo, Yasmiana, additional, Pérez Sánchez, Lincidio, additional, Hernández-Bernal, Francisco, additional, Mendoza Fuentes, Osmany, additional, Selman-Housein, Katty-Hind, additional, Gavilondo Cowley, Jorge Víctor, additional, and Ayala Avila, Marta, additional

Published
2016
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8. Proteomic Study to Survey the CIGB-552 Antitumor Effect

Author

Rodríguez-Ulloa, Arielis, primary, Gil, Jeovanis, additional, Ramos, Yassel, additional, Hernández-Álvarez, Lilian, additional, Flores, Lisandra, additional, Oliva, Brizaida, additional, García, Dayana, additional, Sánchez-Puente, Aniel, additional, Musacchio-Lasa, Alexis, additional, Fernández-de-Cossio, Jorge, additional, Padrón, Gabriel, additional, González López, Luis J., additional, Besada, Vladimir, additional, and Guerra-Vallespí, Maribel, additional

Published
2015
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9. Improved production of the recombinant phospholipase A1 from Polybia paulistawasp venom expressed in bacterial cells for use in routine diagnostics

Author

Perez-Riverol, Amilcar, Musacchio-Lasa, Alexis, Fernandes, Luis Gustavo Romani, dos Santos-Pinto, Jose Roberto Aparecido, Esteves, Franciele Grego, Bazon, Murilo Luiz, Zollner, Ricardo de Lima, Palma, Mario Sergio, and Brochetto-Braga, Márcia Regina

Abstract

Phospholipase A1 (PLA1) is one of the three major allergens identified in the venom of P. paulista(Hymenoptera: Vespidae), a clinically relevant wasp from southeastern Brazil. The recombinant form of this allergen (rPoly p 1) could be used for the development of molecular diagnostic of venom allergy. Early attempts to produce rPoly p 1 using Escherichia coliBL21 (DE3) cells rendered high yields of the insoluble rPoly p 1 but with low levels of solubilized protein recovery (12%). Here, we aimed to improve the production of rPoly p 1 in E. coliby testing different conditions of expression, solubilization of the inclusion bodies and protein purification. The results showed that the expression at 16 °C and 0.1 mM of IPTG increased the production of rPoly p 1, still in the insoluble form, but with high solubilized protein yields after incubation with citrate–phosphate buffer with 0.15 M NaCl, 6 M urea, pH 2.6 at 25 ºC for 2 h. The venom allergen was also cloned in pPICZαA vector for soluble expression as a secreted protein in Pichia pastorisX-33 cells, rendering almost undetectable levels (nanograms) in the culture supernatant. In contrast, a sevenfold increase of the solubilized and purified rPoly p 1 yields (1.5 g/L of fermentation broth) was obtained after improved production in E. coli. The identity of the protein was confirmed with an anti-His antibody and MS spectra. Allergen-specific IgE (sIgE)-mediated recognition was evaluated in immunoblotting with sera of allergic patients (n= 40). Moreover, rPoly p 1 showed high levels of diagnostic sensitivity (95%). The optimized strategy for rPoly p 1 production described here, will provide the amounts of allergen necessary for the subsequent protein refolding, immunological characterization steps, and ultimately, to the development of molecular diagnostic for P. paulistavenom allergy.

Published
2020
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10. Novel molecular events related to CIGB-300 antineoplastic mechanism of action.

Author

Perea Rodríguez, Silvio Ernesto, Perera Negrin, Yasser, Rodríguez Ulloa, Arielis, Ramos Gómez, Yassel, Rosales Menzoney, Mauro, Padrón Palomares, Gabriel, Caballero Menéndez, Evelin, Guirola Cruz, Osmany, Musacchio Lasa, Alexis, Fernández De Cossio, Jorge, González López, Luis Javier, Besada López, Vladimir, Pérez Viltre, George V., Aguilar Noriega, Daylen, Vázquez Blomquist, Dania Marcia, and Ramón Sánchez, Ailyn de la Caridad

Subjects
*PEPTIDE drugs, *WESTERN immunoblotting, *ANTINEOPLASTIC agents, *PROTEIN kinase CK2, *LUNG cancer, *PROTEOMICS, *DRUG synergism, *PEPTIDES, *CISPLATIN
Abstract

CIGB-300 is an antitumor peptide that inhibits the CK2-mediated phosphorylation by direct targeting of substrates. This paper aims to describe breakthroughs about the CIGB-300 antineoplastic mechanism related to the inhibition of CK2-mediated phosphorylation and the synergism with anticancer drugs. CK2 phosphorylation assays were performed with catalytic CK2α subunit or the CK2 holoenzyme in presence or not of CIGB-300. CIGB-300/CK2 interaction was verified by pull-down experiments, in situ colocalization and phosphoproteomic analysis of CIGB-300-treated lung cancer cells were also performed. Synergism of the peptide with anticancer drugs was evaluated in vitro and for Cisplatin; it was also tested in vivo. Besides, comparative proteomics of CIGB-300 combined with Cisplatin was conducted. CIGB-300 targeted the CK2α subunit and inhi-bited the enzymatic activity of the holoenzyme in different experimental settings. Likewise, phosphoproteomic and Western Blot analysis allowed for knowing the early CK2 inhibition profile elicited by CIGB-300 at 10 and 30 min of treatment. Moreover, CIGB-300 did synergize with chemotherapeutics and EGFR inhibitors, and molecular events that reportedly support such synergistic interactions were also known. CIGB-300 inhibits the CK2-mediated phosphorylation by using an alternative mechanism of direct interaction with the enzyme itself. Besides, CIGB-300 synergizes with chemotherapeutics and EGFR inhibitors by modulating different proteins related to drug resistance. [ABSTRACT FROM AUTHOR]

Published
2022

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