Article
Insights into the tracking of the cysteine proteinase B COOH-terminal polypeptide of Leishmania (Leishmania) amazonensis by surface plasmon resonance
Registro en:
SOUZA, Raquel Santos de et al. Insights into the tracking of the cysteine proteinase B COOH-terminal polypeptide of Leishmania (Leishmania) amazonensis by surface plasmon resonance. Parasitology Research, v. 118, p.1249–1259, Feb. 2019.
0932-0113
10.1007/s00436-019-06238-5
1432-1955
Autor
Souza, Raquel Santos de
Silva, Franklin Souza
Melo, Barbara Cristina de Albuquerque
Guimarães, Michelle Lopes Ribeiro
Côrtes, Luzia Monteiro de Castro
Pereira, Bernardo Acácio Santini
Almeida, Mariana Silva
Finkelstein, Léa Cysne
Oliveira Junior, Francisco Odencio Rodrigues de
Pereira, Mirian Claudia de Souza
Alves, Carlos Roberto
Resumen
Leishmania (Leishmania) amazonensis has adaptive mechanisms to the host environment that are guided by its proteinases, including cysteine proteinase B (CPB), and primarily its COOH-terminal region (Cyspep). This work aimed to track the fate of Cyspep by surface plasmon resonance (SPR) of promastigotes and amastigotes to gain a greater understanding of the adaptation of this parasite in both hosts. This strategy consisted of antibody immobilization on a COOH1 surface, followed by interaction with parasite proteins and epoxysuccinyl-L-leucylamido(4-guanidino)butane (E-64). Pro-CPB and Cyspep were detected using specific polyclonal antibodies against a recombinant Cyspep in both parasite forms. The parasitic supernatants from amastigotes and promastigotes exhibited higher anti-Cyspep recognition compared with that in the subcellular fractions. As the supernatant of the promastigote cultures exhibited resonance unit values indicative of an effective with to E-64, this result was assumed to be Pro-CPB detection. Finally, after using three sequential SPR assay steps, we propose that amastigotes and promastigotes release Cyspep into the extracellular environment, but only promastigotes release this polypeptide as Pro-CPB. 2022-01-01