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Increasing in Cysteine Proteinase B Expression and Enzymatic Activity During in Vitro Differentiation of Leishmania (Viannia) Braziliensis: First Evidence of Modulation During Morphological Transition

Increasing in Cysteine Proteinase B Expression and Enzymatic Activity During in Vitro Differentiation of Leishmania (Viannia) Braziliensis: First Evidence of Modulation During Morphological Transition

Cinthia Bernardes Gomes, Franklin Souza -Silva, Karen Dos Santos Charret, Bernardo Acácio Santini Pereira, Léa Cysne Finkelstein, Raquel Santos-de-Souza, Luzia Monteiro de Castro Côrtes, Mirian Claudia Souza Pereira, etc.

Biochimie. 2017 Feb;133:28-36.

PMID: 27919786

Abstract:

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	CAS Number	Price
AP65147220	Z-Phe-Arg 7-amido-4-methylcoumarin hydrochloride	65147-22-0	Price
AP70375223	Z-L-Arg 7-amido-4-methylcoumarin hydrochloride	70375-22-3	Price
AP957311374	Fmoc-L-glutamic acid 1-(7-amido-4-methylcoumarin)	957311-37-4	Price
IAR4241379	Z-Phe-Phe-fluoromethyl ketone		Price

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