Increasing in cysteine proteinase B expression and enzymatic activity during in vitro differentiation of Leishmania (Viannia) braziliensis: First evidence of modulation during morphological transition

• Published in: Biochimie 2017-02, Vol.133, p.28-36
• Main Authors: Gomes, Cinthia Bernardes, -Silva, Franklin Souza, Charret, Karen dos Santos, Pereira, Bernardo Acácio Santini, Finkelstein, Léa Cysne, Santos-de-Souza, Raquel, de Castro Côrtes, Luzia Monteiro, Pereira, Mirian Claudia Souza, Rodrigues de Oliveira, Francisco Odêncio, Alves, Carlos Roberto
• Format: Article
• Language: English
• Subjects: Animals; Cathepsin L; Cathepsins - biosynthesis; Cathepsins - genetics; Cathepsins - metabolism; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cysteine Proteases - biosynthesis; Cysteine Proteases - genetics; Cysteine Proteases - metabolism; Cysteine Proteinase Inhibitors - pharmacology; Cysteine proteinases B; Gene Expression Regulation, Enzymologic - drug effects; Hydrogen-Ion Concentration; Leishmania (Viannia) braziliensis; Leishmania braziliensis - enzymology; Leishmania braziliensis - growth & development; Proteolysis - drug effects; Temperature; Trans-epoxysuccinyl-l-leucylamido(4-guanidino)butane; Z-Phe-Arg 7-amido-4-methylcoumarin hydrochloride; Z-Phe-Phe-fluoromethyl ketoneand
• ISSN: 0300-9084; 1638-6183
• DOI: 10.1016/j.biochi.2016.11.015

Leishmania (Viannia) braziliensis presents adaptive protease-dependent mechanisms, as cysteine proteinases B (CPB). This study investigates the expression of three cpb gene isoforms and CPB enzymatic activity during the parasite differentiation. Relative expression levels of LbrM.08.0810 gene were assessed, exhibiting a higher quantity of transcripts in the logarithmic promastigotes phase than in the stationary promastigotes phase (>1.5 times). The cbp gene tends to decrease during acid pH shock and increases when the temperature rises (>1.3 times). LbrM.08.0820 and LbrM.08.0830 genes exhibited similar expression profiles to LbrM.08.0810 gene, with lower levels being observed overall. The proteolytic activity exhibits a gradual increase during the parasite's differentiation with low levels in samples of logarithmic promastigotes phase (3.2 ± 0.08 mmol min−1 mg protein−1) to a peak of activity after 72 h of incubation at 32 °C (4.2 ± 0.026 mmol min−1 mg protein−1) followed by a subsequent decrease of 68 % of peak activity levels after 96 h of incubation at 32 °C (2.8 ± 0.37 mmol min−1 mg protein−1). These activities were also measured in the presence of selective inhibitors for cysteine proteinases, such as Z-Phe-Phe-fluoromethyl ketone and trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane, demonstrating their source as cathepsin-like proteinases. To the best of our knowledge, this report presents the first description of a modulation of cathepsin L-like expression during the L. (V.) braziliensis in vitro differentiation induced by acid pH and high temperature. •Increase of cathepsin L-like during in vitro differentiation of L. (V.) braziliensis.•Modulation of cathepsin L-like in L. (V.) braziliensis by pH and temperature.•LbrM.08.0820/LbrM.08.0830 genes present expression profiles similar to LbrM.08.0810•Proteolytic activity is increased along differentiation by cathepsin-like.