Toxocara canis: Molecular cloning, characterization, expression and comparison of the kinetics of cDNA-derived arginine kinase

Wickramasinghe, Susiji; Uda, Kouji; Nagataki, Mitsuru; Yatawara, Lalani; Rajapakse, R. P. V. J.; Watanabe, Yuji; Suzuki, Tomohiko; Agatsuma, Takeshi

doi:10.1016/j.exppara.2007.03.015

Cited by 24 publications

(23 citation statements)

References 43 publications

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“…Further, we estimated the kinetics of this enzyme such as K m , K d , K cat , and V max and compared it with other phosphagen kinases (Wickramasinghe et al 2007). …”

Section: Resultsmentioning

confidence: 99%

“…Amplification of Toxocara canis AK using KOD-plus DNA polymerase Toxocara canis AK was amplified as previously described method (Wickramasinghe et al 2007). Briefly, polymerase chain reactions (PCRs) were performed in a total volume of 50 μl.…”

Section: Methodsmentioning

confidence: 99%

“…In vertebrates, creatine kinase (CK) is the only phosphagen kinase and its corresponding substrate is phosphocreatine (Watts 1968(Watts , 1975van Thoai 1968;Morrison 1973). Recently, we reported the molecular cloning, characterization, expression, and comparison of the kinetics of complementary DNA (cDNA)-derived arginine kinase of T. canis (Wickramasinghe et al 2007). As this enzyme is not present in mammals, it might be a useful diagnostic marker for VLM.…”

Section: Introductionmentioning

confidence: 99%

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Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model

et al. 2008

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Toxocariasis is a worldwide zoonotic disease caused by the ascarid nematode Toxocara canis. The most common method available for serodiagnosis of toxocariasis is an enzyme-linked immunosorbent assay (ELISA) test using Toxocara excretory-secretory antigen (TES). The present study describes the development of IgG-ELISA based on antiserum prepared against the recombinant arginine kinase of Toxocara canis. Antiserum was prepared against the purified recombinant arginine kinase (AK) using 6-week-old female Japanese white rabbits. Serum samples were collected from experimentally infected BALB/c and C57BL/6 mice at different time periods. The IgG-ELISA was performed using serum samples from mice (infected/uninfected) and TES antigen with antiserum prepared against the recombinant-AK. The optical density (OD450) was measured at 450 nm using a micro-plate ELISA reader. There were significant differences (P<0.01) in the absorbance between infected and control serum samples. Further, we obtained 100% sensitivity for the serum samples from T. canis-infected mice. Therefore, it is suggested that the recombinant-AK based IgG-ELISA could be applied for immunodiagnosis of human toxocariasis. However, it is necessary to evaluate the specificity of this recombinant antigen with similar geohelminth infections.

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“…Further, we estimated the kinetics of this enzyme such as K m , K d , K cat , and V max and compared it with other phosphagen kinases (Wickramasinghe et al 2007). …”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model

et al. 2008

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“…Phosphagens are phosphorylated guanidine compounds that are linked to ATP by way of a reversible reaction catalyzed by phosphagen (guanidine) kinases (phosphagen+MgADP +H + ↔ guanidine acceptor+MgATP) (Wickramasinghe et al 2007). The traditional and conventional view is that phosphagens function as ATP buffers, permitting maintenance of high ATP values during periods in which there is disequilibrium of ATP supply and demand (Kammermeier 1987(Kammermeier , 1993.…”

Section: Introductionmentioning

confidence: 99%

Immunolocalization of arginine kinase (AK) in Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides

et al. 2012

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Arginine kinase (AK) is a member of the phosphagen kinase family. AK plays a major role in cellular energy metabolism in invertebrates including nematodes. In the present study, we performed the direct immunofluorescence test to determine the immunolocalization of AK in different stages of the life cycle (eggs, larvae, and adult worms) of Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides. Our results indicated variable levels of expression of AK in different stages. Moreover, strong fluorescence was observed in cleaving eggs than in dormant eggs. The highest activity of the enzyme was observed in the fully developed eggs. This may be due to high expression of AK in embryonic development, which is associated with increased energy demand due to cleavage and cellular differentiation. Surprisingly, expression of AK is significantly higher in the middle part and posterior end compared to anterior end of the larvae. In addition, AK is highly concentrated in cellular and metabolically active parts of the body such as hypodermis, muscle, intestine, ovaries, oviducts, and uterus, while it is absent in noncellular areas like cuticle. The present study revealed the presence of AK in T. canis, A. lumbricoides, and T. vitulorum and that it plays a major role in energy metabolism of these nematodes. Interestingly, antiserum was prepared against the recombinant T. canis AK and reacts with the native AKs of T. canis, A. lumbricoides, and T. vitulorum. AK levels could vary in relation to maximum potential rates of ATP turnover, oxidative capacity, and energy output. Further studies on subcellular localization of AK in these important helminths provide new information for researchers to develop effective anthelmintics against the parasites of veterinary and of public health importance.

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“…Besides T. cruzi, phosphagen kinases have also been identified in other important animal and human parasites. AKs were cloned from the nematodes Ascaris suum and Toxocara canis which can cause visceral larva migrans (VLM) in humans (Nagataki et al, 2008;Wickramasinghe et al, 2007) and from T. brucei which causes human sleeping sickness and Nagana in livestock (Pereira et al, 2002). The PK from the trematode Schistosoma mansoni was also recently described by Awama et al (2008).…”

Section: Cloning and Molecular Characterization Of P Westermani Pkmentioning

confidence: 99%

Phosphagen Kinase System of the Trematode Paragonimus westermani: Cloning and Expression of a Novel Chemotherapeutic Target

Jarilla¹,

Agatsuma²

2011

Molecular Cloning - Selected Applications in Medicine and Biology

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Toxocara canis: Molecular cloning, characterization, expression and comparison of the kinetics of cDNA-derived arginine kinase

Cited by 24 publications

References 43 publications

Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model

Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model

Immunolocalization of arginine kinase (AK) in Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides

Phosphagen Kinase System of the Trematode Paragonimus westermani: Cloning and Expression of a Novel Chemotherapeutic Target

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