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Yield improvement and enzymatic dissection of Plasmodium falciparum plasmepsin V
In this study, we employed a multipurpose fusion tag to improve the production of PfPMV in E. coli. Recombinant PfPMVm, comprising residues 84 to 521, was substantially obtained in soluble form and could be purified in a single step, yielding a 3.7-fold increase in purified PfPMVm compared to previous reports. Additionally, we have mutated the catalytic residues (D118 N and D365 N), individually and together, and the unpaired cysteine residue C178 to evaluate the effects on catalytic efficiency. Mutation of D365 had more pronounced effects on the catalytic efficiency than that of D118, suggesting that the D365 may act ...
Source: Molecular and Biochemical Parasitology - May 18, 2019 Category: Parasitology Source Type: research

Characterization of a Schistosoma mansoni NDPK expressed in sexual and digestive organs
Publication date: Available online 16 May 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Juliana Roberta Torini, Adriano de Freitas Fernandes, Vitor Hugo Balasco Serrão, Larissa Romanello, Louise E Bird, Joanne E Nettleship, Raymond J. Owens, José Brandão-Neto, Ana Eliza Zeraik, Ricardo DeMarco, Humberto D'Muniz PereiraAbstractNucleoside diphosphate kinases (NDPKs) are crucial to keep the high triphosphate nucleotide levels in the biological process. The enzymatic mechanism has been extensively described; however, the structural characteristics and kinetic parameters have never been fully determined. In Sc...
Source: Molecular and Biochemical Parasitology - May 17, 2019 Category: Parasitology Source Type: research

Editorial Board
Publication date: June 2019Source: Molecular and Biochemical Parasitology, Volume 230Author(s): (Source: Molecular and Biochemical Parasitology)
Source: Molecular and Biochemical Parasitology - May 15, 2019 Category: Parasitology Source Type: research

Palmitoylation in apicomplexan parasites: from established regulatory roles to putative new functions
Publication date: Available online 9 April 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Maria M. Corvi, Valeria R. TurowskiAbstractThis minireview aims to provide a comprehensive synthesis on protein palmitoylation in apicomplexan parasites and higher eukaryotes where most of the data is available. Apicomplexan parasites encompass numerous obligate intracellular parasites with significant health risk to animals and humans. Protein palmitoylation is a widespread post-translational modification that plays important regulatory roles in several physiological and pathological states. Functional studies demonstra...
Source: Molecular and Biochemical Parasitology - April 10, 2019 Category: Parasitology Source Type: research

Editorial Board
Publication date: April 2019Source: Molecular and Biochemical Parasitology, Volume 229Author(s): (Source: Molecular and Biochemical Parasitology)
Source: Molecular and Biochemical Parasitology - March 29, 2019 Category: Parasitology Source Type: research

Farnesyl pyrophosphate synthase is essential for the promastigote and amastigote stages in Leishmania major
Publication date: Available online 26 March 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Sumit Mukherjee, Somrita Basu, Kai ZhangAbstractIsoprenoid synthesis provides a diverse class of biomolecules including sterols, dolichols, ubiquinones and prenyl groups. The enzyme farnesyl pyrophosphate synthase (FPPS) catalyzes the formation of farnesyl pyrophosphate, a key intermediate for the biosynthesis of all isoprenoids. In Leishmania, FPPS is considered the main target of nitrogen containing bisphosphonates, yet the essentiality of this enzyme remains untested. Using a facilitated knockout approach, we carried...
Source: Molecular and Biochemical Parasitology - March 27, 2019 Category: Parasitology Source Type: research

In-solution behavior and protective potential of asparagine synthetase A from Trypanosoma cruzi
Publication date: Available online 15 March 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Flávia Souza Morini, Emanuella de Castro, Stephanie Bath de Morais, Cesar Lancheros Contreras, Ana Valéria Pereira Weiler, Mario Tyago Murakami, Phileno Pinge-Filho, Sueli Fumie Yamada-Ogatta, Tatiana de Arruda Campos Brasil de SouzaAbstractL-Asparagine synthetase (AS) acts in asparagine formation and can be classified into two families: AS-A or AS-B. AS-A is mainly found in prokaryotes and can synthetize asparagine from ammonia. Distinct from other eukaryotes, Trypanosoma cruzi produces an AS-A. AS-A from Trypanosoma...
Source: Molecular and Biochemical Parasitology - March 17, 2019 Category: Parasitology Source Type: research

Characterization of strain-specific phenotypes associated with knockout of dense granule protein 9 in Toxoplasma gondii
Publication date: Available online 5 March 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Huanping Guo, Yang Gao, Honglin Jia, Paul Franck Adjou Moumouni, Tatsunori Masatani, Mingming Liu, Seung-Hun Lee, Eloiza May Galon, Jixu Li, Yongchang Li, Maria Agnes Tumwebaze, Byamukama Benedicto, Xuenan XuanAbstractToxoplasma gondii is an obligate intracellular parasite that can invade any nucleated cell of mammals and cause toxoplasmosis. Dense granule proteins play major structural functions within the parasitophorous vacuole (PV) and the cyst wall of T. gondii. Moreover, their particular location within the PV allo...
Source: Molecular and Biochemical Parasitology - March 6, 2019 Category: Parasitology Source Type: research

Immunofluorescence staining protocol for STED nanoscopy of Plasmodium-infected red blood cells
Publication date: Available online 1 March 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Ann-Kathrin Mehnert, Caroline Sophie Simon, Julien GuizettiAbstractImmunofluorescence staining is the key technique for visualizing organization of endogenous cellular structures in single cells. Labeling and imaging of blood stage Plasmodium falciparum has always been challenging since it is a small intracellular parasite. A widely-used standard for parasite immunofluorescence is fixation in suspension with addition of minute amounts of glutaraldehyde to the paraformaldehyde-based solution. While this maintains red bloo...
Source: Molecular and Biochemical Parasitology - March 5, 2019 Category: Parasitology Source Type: research

PROTEOMIC ANALYSIS OF GLYCOSOMES FROM Trypanosoma cruzi EPIMASTIGOTES
In this study, we have purified glycosomes from T. cruzi epimastigotes, collected the soluble and membrane fractions of these organelles, and separated peripheral and integral membrane proteins by Na2CO3 treatment and osmotic shock. Proteomic analysis was performed on each of these fractions, allowing us to confirm the presence of enzymes involved in various metabolic pathways as well as identify new components of this parasite’s glycosomes.Graphical abstract (Source: Molecular and Biochemical Parasitology)
Source: Molecular and Biochemical Parasitology - March 5, 2019 Category: Parasitology Source Type: research

Transcriptomic profile of two developmental stages of the cestode parasite Mesocestoides corti
Publication date: Available online 21 February 2019Source: Molecular and Biochemical ParasitologyAuthor(s): T. Basika, G.P. Paludo, F.M. Araujo, A.C. Salim, F. Pais, L. Maldonado, N. Macchiaroli, J. Camargo de Lima, M. Rosenzvit, G. Oliveira, L. Kamenetzky, H.B. FerreiraAbstractCestode development involves complex morphological and physiological changes. Here, we performed a differential expression analysis of gene transcripts between two developmental stages of the model cestode Mesocestoides corti. A RNA-seq-based approach was used to compare the transcriptomes of the tetrathyridium (larval, TT) and strobilated worm (ST)...
Source: Molecular and Biochemical Parasitology - February 22, 2019 Category: Parasitology Source Type: research

Cloning, expression and purification of 3'-nucleotidase/nuclease, an enzyme responsible for the Leishmania escape from neutrophil extracellular traps
This study aims to deepen the knowledge about 3’NT/NU, mainly with respect to its nuclease activity that is little studied in the current literature. For this, we cloned, expressed and purified the recombinant La3'NT/NU and have confirmed its contribution to the parasite escape from NET-mediated killing.Graphical abstract (Source: Molecular and Biochemical Parasitology)
Source: Molecular and Biochemical Parasitology - February 15, 2019 Category: Parasitology Source Type: research

Control and regulation of the pyrophosphate-dependent glucose metabolism in Entamoeba histolytica
Publication date: Available online 14 February 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Emma Saavedra, Rusely Encalada, Citlali Vázquez, Alfonso Olivos-García, Paul A.M. Michels, Rafael Moreno-SánchezAbstractEntamoeba histolytica has neither Krebs cycle nor oxidative phosphorylation activities; therefore, glycolysis is the main pathway for ATP supply and provision of carbon skeleton precursors for the synthesis of macromolecules. Glucose is metabolized through fermentative glycolysis, producing ethanol as its main end-product as well as some acetate. Amoebal glycolysis markedly differs from the typic...
Source: Molecular and Biochemical Parasitology - February 15, 2019 Category: Parasitology Source Type: research

In vitro and in vivo characterization of the multiple isoforms of Schistosoma mansoni Hypoxanthine-guanine phosphoribosyltransferases
Publication date: Available online 14 February 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Larissa Romanello, Ana Eliza Zeraik, Adriano de Freitas Fernandes, Juliana Roberta Torini, Louise E. Bird, Joanne E. Nettleship, Heather Rada, Yamini Reddivari, Ray J. Owens, Vitor Hugo Balasco Serrão, Ricardo DeMarco, José Brandão-Neto, Humberto D'Muniz PereiraAbstractSchistosoma mansoni, the parasite responsible for schistosomiasis, lacks the “de novo” purine biosynthetic pathway and depends entirely on the purine salvage pathway for the supply of purines. Numerous reports of praziquantel resistance have bee...
Source: Molecular and Biochemical Parasitology - February 15, 2019 Category: Parasitology Source Type: research

Nuclear localization signals in trypanosomal proteins
Publication date: Available online 14 February 2019Source: Molecular and Biochemical ParasitologyAuthor(s): Israel Canela-Pérez, Imelda López-Villaseñor, Luis Mendoza, Ana María Cevallos, Roberto HernándezABSTRACTThe nuclear import of proteins in eukaryotic cells is a fundamental biological process. While it has been analysed to different extents in model eukaryotic organisms, this event has rarely been studied in the early divergent protozoa of the order Kinetoplastida. The work presented here represents an overview of nuclear import in these important species of human pathogens. Initially, an in silico study of clas...
Source: Molecular and Biochemical Parasitology - February 15, 2019 Category: Parasitology Source Type: research