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2022-01-01
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- IOC - Artigos de Periódicos [12130]
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INSIGHTS INTO THE TRACKING OF THE CYSTEINE PROTEINASE B COOH-TERMINAL POLYPEPTIDE OF LEISHMANIA (LEISHMANIA) AMAZONENSIS BY SURFACE PLASMON RESONANCE
Cysteine proteinase
Trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane
Surface plasmon resonance
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Abstract
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.
Keywords
Leishmania (L.) amazonensisCysteine proteinase
Trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane
Surface plasmon resonance
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