Leishmania (L.) amazonensis peptidase activities inside the living cells and in their lysates

Molecular and Biochemical Parasitology
Elide E CaroselliLuiz Juliano

Abstract

In this study we investigated the peptidase activity in Leishmania (L.) amazonensis live amastigote by confocal microscopy using peptidyl-MCA as substrates, the hydrolysis of which releases the MCA fluorophore inside the cells. Cell pre-treatment with peptidase inhibitors indicated the presence of cysteine and serine peptidases. It was noteworthy that Leishmania amastigotes incorporate only substrates (Z-FR-MCA, Z-RR-MCA) or inhibitors (E64, TLCK) containing positively charged groups. The peptidase activities in the supernatants of amastigotes and promastigotes lysates were also evaluated with the same peptidyl-MCA substrates and inhibitors in the pH range 4.5-9.0. The effects of temperature and different salts were also included in this study. The hydrolytic activities of supernatants on Z-FR-MCA clearly indicate the presence of different cysteine peptidases that adapted to work in different environment conditions. Intact Leishmania cells incorporated Z-RR-MCA, the hydrolysis of which was inhibited only by TLCK indicating the presence of at least one serine peptidase. The pH profile of Z-RR-MCA hydrolysis by amastigotes and promastigotes lysate supernatants, and the hydrolysis time course of the FRET peptide Abz-AGRRRAQ-EDDnp ...Continue Reading

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Citations

Jun 14, 2016·European Journal of Medicinal Chemistry·Paulo André Teixeira de Moraes GomesAna Cristina Lima Leite
Oct 20, 2019·Biochimie·Flávia Nader MottaIzabela Marques Dourado Bastos
Apr 8, 2019·Experimental Parasitology·José C QuillesAndrei Leitão
Aug 23, 2021·Bioorganic & Medicinal Chemistry·Wagner A S JudiceTiago Rodrigues

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