Characterization of the protein tyrosine phosphatase PRL from Entamoeba histolytica

Ramírez-Tapia, Ana Lilia; Baylón-Pacheco, Lidia; Espíritu-Gordillo, Patricia; Rosales‐Encina, José Luis

doi:10.1016/j.exppara.2015.09.014

Cited by 7 publications

(5 citation statements)

References 56 publications

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“…verum was found to be inhibited by established commercial inhibitors of PTPases (sodium metavanadate) in a dose‐dependent manner, whereas another inhibitor, NaF (Ser/Thr and acidic phosphatase inhibitor) was not effective at low concentrations (5 and 10 mM) but partially inhibited at a concentration of 20 mM (Figure 9). These results are in good agreement with existing studies on EhPRL and TcPRL‐1, in which phosphatase activity was inhibited by sodium o‐vanadate (0.6 mM) and unaffected by 5 mM NaF (Cuevas et al., 2005; Ramírez‐Tapia et al., 2015).…”

Section: Resultssupporting

confidence: 92%

Trachyspermum ammi and Cinnamomum verum as nutraceuticals: Spices rich in therapeutically significant protein tyrosine phosphatases

et al. 2021

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Nutraceuticals need special attention as preventive molecules to create a natural barrier against various dreadful diseases like cancer and to regulate metabolism. In the present study, two spices, Trachyspermum ammi and Cinnamomum verum, been identified as excellent Protein Tyrosine Phosphatases (PTPases) sources that play significant role in the regulation of cell signal transduction and developmental processes in plants as well as animals, being lucrative and potential targets for pharmacological modulation. PTPases from both cases were partially purified into 0%–40% and 40%–80% fractions based on ammonium sulfate saturation levels. Fraction (40%–80%) exhibited a purification level of 4.44‐fold and 2.86‐fold with specific activity of 44.06 and 23.33 U/mg for PTPases from T. ammi and C. verum, respectively. PTPases being found to be thermally stable up to 70°C imply their industrial significance. Kinetic studies showed Km values to be 7.14 and 8.33 mM, whereas the activation energy (Ea) values were 25.89 and 29.13 kJ/mol, respectively. Divalent cations: Cu2+, Zn2+, and Mn2+ acted as inhibitors of PTPases, from both sources. The Ki values of inhibitors varied from 0.014–0.125 mM in the descending order Cu2+ > Zn2+ > Mn2+ and Mn2+ > Cu2+ > Zn2+ for PTPases from T. ammi and C. verum, respectively. The inhibitory effect of sodium metavanadate aligns with prominent PTPase characteristics. In addition to these properties, the thermostability of PTPases from two spices enhances their significance in industries with therapeutically vital products. Although the source of PTPases is culinary spices, further studies are required to establish the utilization of PTPases as nutraceuticals and in therapeutic formulations. Practical applications For a healthy lifestyle, awareness needs to be created by humankind towards food habits to minimize illnesses. Numerous studies have explored the consumption of nutraceutical products acts as a natural barrier and immune booster for various human ailments including SARS‐COV‐2. PTPases play important roles in regulating intracellular signaling and, ultimately, biological function along with their structural features. The importance of PTPases and their inhibitors has been implicated in various diseases like cancer, diabetes, and obesity. Further investigations need to be undertaken to explore the therapeutic properties of PTPases in both in vivo and in vitro for their clinical significance.

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Section: Resultssupporting

confidence: 92%

Trachyspermum ammi and Cinnamomum verum as nutraceuticals: Spices rich in therapeutically significant protein tyrosine phosphatases

et al. 2021

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“…Both these genes are differentially expressed in trophozoites from amoebic liver abscess [ 22 ]. E. histolytica also has a PRL phosphatase (a type of PTP) located in small patches on the outer membrane and in internal vesicles and is known to dephosphorylate tyrosine residues [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

Protein Phosphatase PP2C Identification in Entamoeba spp

Navarrete-Mena

Pacheco‐Yépez

Hernández‐Ramírez

et al. 2021

BioMed Research International

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Entamoeba histolytica is the causative agent of amoebiasis, and Entamoeba dispar is its noninvasive morphological twin. Entamoeba invadens is a reptilian parasite. In the present study, Western blot, phosphatase activity, immunofluorescence, and bioinformatic analyses were used to identify PP2C phosphatases of E. histolytica, E. dispar, and E. invadens. PP2C was identified in trophozoites of all Entamoeba species and cysts of E. invadens. Immunoblotting using a Leishmania mexicana anti-PP2C antibody recognized a 45.2 kDa PP2C in all species. In E. histolytica and E. invadens, a high molecular weight element PP2C at 75 kDa was recognized, mainly in cysts of E. invadens. Immunofluorescence demonstrated the presence of PP2C in membrane and vesicular structures in the cytosol of all species analyzed. The ~75 kDa PP2C of Entamoeba spp. shows the conserved domain characteristic of phosphatase enzymes (according to in silico analysis). Possible PP2C participation in the encystation process was discussed.

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“…ECM components have been used to coat surfaces where trophozoites are grown, for example collagen and FN (Ramírez-Tapia et al, 2015; Talamás-Lara et al, 2015), and these interaction trophozoite-substrate leads to a response as the differential expression of proteins, protease secretion (Piña-Vázquez et al, 2012), and cytoskeletal rearrangements in which actin polymerization is actively involved (Sengupta et al, 2009; Javier-Reyna et al, 2012; Starke et al, 2014; Aguilar-Rojas et al, 2016). EMC components (such as FN) have been used to generate micropatterns in order to stimulate eukaryotic cells to migrate, produce abundant and different membrane protrusions (Alamdari et al, 2013; Starke et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…E. histolytica trophozoites, strain HM1:IMSS (ATCC 30459), clone A (Ramírez-Tapia et al, 2015), were axenically cultured at 37°C in TYI-S-33 medium supplemented with 10% heat-inactivated adult bovine serum (ABS) and harvested during logarithmic growth phase (Diamond et al, 1978). All experiments presented here were performed on at least three separate occasions and in triplicate.…”

Section: Methodsmentioning

confidence: 99%

Influence of Micropatterned Grill Lines on Entamoeba histolytica Trophozoites Morphology and Migration

Sierra-López

Baylón-Pacheco

Espíritu-Gordillo

et al. 2018

Front. Cell. Infect. Microbiol.

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Entamoeba histolytica, the causal agent of human amoebiasis, has two morphologically different phases: a resistant cyst and a trophozoite responsible for the invasion of the host tissues such as the colonic mucosa and the intestinal epithelium. During in vitro migration, trophozoites usually produce protuberances such as pseudopods and rarely filopodia, structures that have been observed in the interaction of trophozoites with human colonic epithelial tissue. To study the different membrane projections produced by the trophozoites, including pseudopods, filopodia, uropods, blebs, and others, we designed an induction system using erythrocyte extract or fibronectin (FN) in micropatterned grill lines (each micro-line containing multiple micro-portions of FN or erythrocyte extract) on which the trophozoites were placed in culture for migration assays. Using light, confocal, and scanning electron microscopy, we established that E. histolytica trophozoites frequently produce short and long filopodia, large retractile uropods in the rear, pseudopods, blebs, and others structures, also showing continuous migration periods. The present study provides a simple migration method to induce trophozoites to generate abundant membrane protrusion structures that are rarely obtained in normal or induced cultures, such as long filopodia; this method will allow a–better understanding of the interactions of trophozoites with FN and cell debris. E. histolytica trophozoites motility plays an important role in invasive amoebiasis. It has been proposed that both physical forces and chemical signals are involved in the trophozoite motility and migration. However, the in vivo molecules that drive the chemotactic migration remain to be determined. We propose the present assay to study host molecules that guide chemotactic behavior because the method is highly reproducible, and a live image of cell movement and migration can be quantified.

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Characterization of the protein tyrosine phosphatase PRL from Entamoeba histolytica

Cited by 7 publications

References 56 publications

Trachyspermum ammi and Cinnamomum verum as nutraceuticals: Spices rich in therapeutically significant protein tyrosine phosphatases

Trachyspermum ammi and Cinnamomum verum as nutraceuticals: Spices rich in therapeutically significant protein tyrosine phosphatases

Protein Phosphatase PP2C Identification in Entamoeba spp

Influence of Micropatterned Grill Lines on Entamoeba histolytica Trophozoites Morphology and Migration

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