Molecular cloning, expression and characterization of a functional GSTmu class from the cattle tick Boophilus annulatus

Shahein, Yasser E.; El-Hakim, Amr E.; Abouelella, Amira M.; Hamed, Ragaa R.; Allam, Shaimaa A.; Farid, Nevin M.

doi:10.1016/j.vetpar.2007.12.014

Cited by 25 publications

(14 citation statements)

References 44 publications

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“…To the best of our knowledge, this is the first report of myconanotechnology originating from the fungus D. flagrans (AC001) as a nematicidal "tool". It should be emphasized that the resistance of A. caninum to conventional drugs is already well known, 29 and results like those presented in this study may help make control more effective.…”

Section: Resultsmentioning

confidence: 53%

Nematicidal activity of silver nanoparticles from the fungus Duddingtonia flagrans

Barbosa¹,

Silva²,

Ferraz³

et al. 2019

IJN

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Background: Helminth parasites cause morbidity and mortality in both humans and animals. Most anthelmintic drugs used in the treatment of parasitic nematode infections act on target proteins or regulate the electrical activity of neurons and muscles. In this way, it can lead to paralysis, starvation, immune attack, and expulsion of the worm. However, current anthelmintics have some limitations that include a limited spectrum of activity across species and the threat of drug resistance, which highlights the need for new drugs for human and veterinary medicine. Purpose: Present study has been conducted to determine the anthelmintic activity of silver nanoparticles (AgNPs) synthesized from the extract of nematophagous fungus, Duddingtonia flagrans, on the infecting larvae of Ancylostoma caninum (L 3 ). Methods: The nanoparticles were characterized by visual, ultraviolet, Fourier-transform infrared spectroscopy, transmission electron microscopy (TEM) analysis, and X-ray diffraction. The in vitro study was based on experiments to inhibit the motility of infective larvae (L 3 ), and the ultrastructural analysis of the nematode was performed by images obtained by TEM. Results: The XRD studies revealed the crystalline nature of the nanoparticles, and FTIR results implied that AgNPs were successfully synthesized and capped with compounds present in the extract. The results showed that the green synthesis of AgNPs exhibited nematicidal activity, being the only ones capable of penetrating the cuticle of the larvae, causing changes in the tegmentum, and consequently, the death of the nematode. Conclusion: The extract of the fungus D. flagrans is able to synthesize AgNP and these have a nematicidal action.

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Section: Resultsmentioning

confidence: 53%

Nematicidal activity of silver nanoparticles from the fungus Duddingtonia flagrans

Barbosa¹,

Silva²,

Ferraz³

et al. 2019

IJN

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“…(B.) annulatus and the active recombinant BaGST was expressed and purified as fusion protein His-tagged protein (Shahein et al, 2008). This GST was later shown to be effectively inhibited by phenolic compounds and flavonoids from plant extracts but also by a haematin (Guneidy et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Inducible glutathione S-transferase (IrGST1) from the tick Ixodes ricinus is a haem-binding protein

Perner

Kotál

Hatalová

et al. 2018

Insect Biochemistry and Molecular Biology

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Blood-feeding parasites are inadvertently exposed to high doses of potentially cytotoxic haem liberated upon host blood digestion. Detoxification of free haem is a special challenge for ticks, which digest haemoglobin intracellularly. Ticks lack a haem catabolic mechanism, mediated by haem oxygenase, and need to dispose of vast majority of acquired haem via its accumulation in haemosomes. The knowledge of individual molecules involved in the maintenance of haem homeostasis in ticks is still rather limited. RNA-seq analyses of the Ixodes ricinus midguts from blood- and serum-fed females identified an abundant transcript of glutathione S-transferase (gst) to be substantially up-regulated in the presence of red blood cells in the diet. Here, we have determined the full sequence of this encoding gene, ir-gst1, and found that it is homologous to the delta-/epsilon-class of GSTs. Phylogenetic analyses across related chelicerates revealed that only one clear IrGST1 orthologue could be found in each available transcriptome from hard and soft ticks. These orthologues create a well-supported clade clearly separated from other ticks' or mites' delta-/epsilon-class GSTs and most likely evolved as an adaptation to tick blood-feeding life style. We have confirmed that IrGST1 expression is induced by dietary haem(oglobin), and not by iron or other components of host blood. Kinetic properties of recombinant IrGST1 were evaluated by model and natural GST substrates. The enzyme was also shown to bind haemin in vitro as evidenced by inhibition assay, VIS spectrophotometry, gel filtration, and affinity chromatography. In the native state, IrGST1 forms a dimer which further polymerises upon binding of excessive amount of haemin molecules. Due to susceptibility of ticks to haem as a signalling molecule, we speculate that the expression of IrGST1 in tick midgut functions as intracellular buffer of labile haem pool to ameliorate its cytotoxic effects upon haemoglobin intracellular hydrolysis.

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“…Whole tick, salivary, larval and gut antigens of R. annulatus were prepared according to the methods described by Shahein et al [22]. In brief, laboratory reared, clean, 5-6-day-old unfed ticks, larvae, salivary glands or midgut isolated by dissection from adult ticks were homogenized in cold buffer A which includes, 0.15 M phosphate-buffered saline (PBS), 1 mM disodium EDTA, pH 7.2, and protease inhibitors cocktail (Sigma-Aldrich, Saint Louis, MO, USA).…”

Section: Preparation Of Whole Tick Larval Salivary and Gut Antigensmentioning

confidence: 99%

“…Briefly, a bacterial culture of BL21 (DE3) transformed with recombinant pET30b-Ra-sHSPII was induced using 100 mM IPTG at OD 0.9-1.1 for 2 h. After centrifugation, the bacterial pellet was washed twice and then resuspended in buffer B (25 mM Tris-base, 10% glycerol, 2 mM DTT and 1 mM EDTA). The same protocol was followed for BL21(DE3) E. coli culture transformed with recombinant pET30b-BaGST [22], as a possible control expressing a protein that does not confer thermal stability.…”

Section: Thermal Stabilitymentioning

confidence: 99%

Molecular cloning of a small heat shock protein (sHSPII) from the cattle tick Rhipicephalus (Boophilus) annulatus salivary gland

Shahein¹,

El-Rahim²,

Hussein³

et al. 2010

International Journal of Biological Macromolecules

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Molecular cloning, expression and characterization of a functional GSTmu class from the cattle tick Boophilus annulatus

Cited by 25 publications

References 44 publications

<p>Nematicidal activity of silver nanoparticles from the fungus <em>Duddingtonia flagrans</em></p>

<p>Nematicidal activity of silver nanoparticles from the fungus <em>Duddingtonia flagrans</em></p>

Inducible glutathione S-transferase (IrGST1) from the tick Ixodes ricinus is a haem-binding protein

Molecular cloning of a small heat shock protein (sHSPII) from the cattle tick Rhipicephalus (Boophilus) annulatus salivary gland

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