Understanding serine proteases implications on Leishmania spp lifecycle

Alves, Carlos Roberto; Souza, Raquel Santos; Charret, Karen dos Santos; Côrtes, Luzia Monteiro de Castro; Sá-Silva, Matheus Pereira de; Barral-Veloso, Laura; Oliveira, Luiz Filipe Gonçalves de; Souza-Silva, Franklin

doi:10.1016/j.exppara.2017.11.008

Cited by 28 publications

(25 citation statements)

References 97 publications

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“…resistance towards antimony treatment is an increasing multifactorial phenomenon, and several virulence factors have been studied and related to resistant phenotypes in clinical isolates [39,[61][62][63]. The present study aids to this discussion by bringing proteases to focus, one of the most studied parasite virulence factors in the last 30 years [18,22,64]. The data presented here shows the possibility to use serine proteases to in vitro characterize and distinguish L. (V.) braziliensis clinical isolates with different responses towards antimony.…”

Section: Discussionmentioning

confidence: 65%

“…In addition, based on the previously role seen for subtilisins as maturases of TXNPx [18,38] TXNPx are expressed by L. (V.) braziliensis clinical isolates, but its level of expression varies depending on the isolate phenotype. Interestingly, a proteomic study using lab-generated L. (V.) braziliensis NR/resistant strains showed that TXNPx abundance was signi cantly higher than R/susceptible strains [60].…”

Section: Discussionmentioning

confidence: 99%

“…Speci cally, serine proteases have been extensively studied in American Leishmania spp. due to their functional interrelationship in parasite physiology and their potential as therapeutic targets [18][19][20][21][22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

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Serine Proteases Profiles of Leishmania (Viannia) Braziliensis Clinical Isolates With Distinct Susceptibilities to Antimony

Zabala-Peñafiel¹,

Dias-Lopes²,

Cysne-Finkelstein

et al. 2020

Preprint

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Background: Glucantime® (SbV) is considered the first-line treatment against American Tegumentary Leishmaniasis in South America, though increased parasite resistance towards it have been reported and hampered its effectiveness. In this context, subtilisins serine proteases have been related to parasites’ susceptibility to drugs such as SbV and derivatives, through modulation of the parasite detoxification system. However, little is known about parasites causing ATL and their distinct responses towards this treatment.Methods: The study was conducted using Leishmania (Viannia) braziliensis clinical isolates from patients that presented clinical cure or Responders (R) and relapse/therapeutic failure or Non-responders (NR). Twelve clinical isolates were used to assess their in vitro susceptibility to SbIII and SbV, serine proteases activity, and expression of subtilisins-like and tryparedoxin-peroxidase (TXNPx) transcripts. Results: SbIII was able to better distinguish axenic amastigotes from each clinical group. These isolates were also assessed for serine protease activity, using z-FR-AMC as substrate and detecting distinct enzyme profiles with specific inhibitors. TLCK inhibited almost 100% of activity in both promastigotes and axenic amastigotes while AEBSF inhibited around 70%. PMSF showed low inhibition of specific isolates (35%). Gathering all the quantitative data, we performed principal component analysis and then used the K-means algorithm to cluster the isolates. This analysis yielded one cluster with only one isolate (R isolate), one homogeneous cluster (NR isolates), and three heterogeneous clusters (R and NR isolates). Additionally, gene transcripts of subtilisins and TXNPx were detected in promastigotes and axenic amastigotes from both groups. Conclusions: Cluster analysis showed that there is a phenotypic heterogeneity among the isolates, however, exploration of in vitro phenotypes based on SbIII and serine proteases profiles can aid in the characterization and better understanding of L. (V.) braziliensis clinical isolates.

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Section: Discussionmentioning

confidence: 65%

Section: Discussionmentioning

confidence: 99%

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Serine Proteases Profiles of Leishmania (Viannia) Braziliensis Clinical Isolates With Distinct Susceptibilities to Antimony

Zabala-Peñafiel¹,

Dias-Lopes²,

Cysne-Finkelstein

et al. 2020

Preprint

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“…The biological cycles of different Leishmania spp. are strongly driven by nuances in their metabolic profiles that are related to the composition of their specific degradomes, including the protease and its respective substrate repertoire [7]. Their enzyme class (EC) is hydrolases (3) subclassified as peptidases (3.4) according to their physicochemical, biochemical, and structural features.…”

Section: Introductionmentioning

confidence: 99%

“…Of the 17 predicted biological functions related to Leishmania sp. serine proteinase genes, only 18% were related to parasite physiology, including their activity as signal peptidases for removing the signal peptide from secretory preproteins, as maturases of other proteins, and as metacaspases [7]. The 26 to 28 serine proteinase genes from Leishmania sp.…”

Section: Introductionmentioning

confidence: 99%

Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression

Santos-de-Souza

Côrtes

Charret

et al. 2019

IJMS

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Serine proteinases in Leishmania (Viannia) braziliensis promastigotes were assessed in this work. This study included the investigation of the enzymatic activity of subcellular fractions obtained from benzamidine affinity chromatography, reverse transcription polymerase chain reactions, and in silico assays of subcellular localization of subtilisin. Promastigote serine proteinases showed gelatinolytic activity with molecular masses of 43 kDa to 170 kDa in the cytosolic fraction and 67 kDa to 170 kDa in the membranous fraction. Serine proteinase activities were detected using N-benzyloxycarbonyl-l-phenylalanyl-l-arginine 7-amino-4-methylcoumarin (Z-FR-AMC) and N-succinyl-l-alanine-l-phenylalanine-l-lysine 7-amino-4-methylcoumarin (Suc-AFK-AMC) as substrates in the cytosolic fraction (Z-FR-AMC = 392 ± 30 µmol.min−1 mg of protein−1 and Suc-AFK-AMC = 252 ± 20 µmol.min−1 mg of protein−1) and in the membranous fraction (Z-FR-AMC = 53 ± 5 µmol.min−1 mg of protein−1 and Suc-AFK-AMC = 63.6 ± 6.5 µmol.min−1 mg of protein−1). Enzyme specificity was shown by inhibition with aprotinin (19% to 80% inhibition) and phenylmethanesulfonyl fluoride (3% to 69%), depending on the subcellular fraction and substrate. The expression of subtilisin (LbrM.13.0860 and LbrM.28.2570) and tryparedoxin peroxidase (LbrM.15.1080) genes was observed by the detection of RNA transcripts 200 bp, 162 bp, and 166 bp long, respectively. Subsequent in silico assays showed LbrM.13.0860 can be located in the cytosol and LbrM.28.2570 in the membrane of the parasite. Data obtained here show the subcellular distribution and expression of serine proteinases, including the subtilisin-like serine proteinases in L. (V.) braziliensis promastigotes.

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Drug Discovery and Development for Kinetoplastid Diseases

Caffrey

Steverding

Ferreira

et al. 2021

Burger's Medicinal Chemistry and Drug Discovery

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In this article, we review the disease, biology, and biochemistry of kinetoplastids, as well as the new drugs and drug candidates that have entered the clinic in the last decade. We also describe examples of the preclinical exploration of small molecules against various protein targets (e.g. cysteine proteases, the proteasome, and tubulin), as well as cutting‐edge molecular and computational strategies and technologies being brought to bear to discover and develop new antitrypanosomal drugs. For comprehensive descriptions of the disease, biology, and drug therapies prior to 2011, the reader is encouraged to review the article by P.M. Woster that appeared in 2010 in the seventh edition of Burger's Medicinal Chemistry, Drug Discovery, and Development, entitled Antiprotozoal/Antiparasitic Agents.

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Understanding serine proteases implications on Leishmania spp lifecycle

Cited by 28 publications

References 97 publications

Serine Proteases Profiles of Leishmania (Viannia) Braziliensis Clinical Isolates With Distinct Susceptibilities to Antimony

Serine Proteases Profiles of Leishmania (Viannia) Braziliensis Clinical Isolates With Distinct Susceptibilities to Antimony

Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression

Drug Discovery and Development for Kinetoplastid Diseases

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