Divalent cation hinder the solubilization of a tubulin kinase activity from Trypanosoma cruzi epimastigotes

Uzcanga, Graciela L.; Galán-Caridad, José M.; Noris-Suárez, Karem; Bubis, José

doi:10.4067/s0716-97602003000300008

Cited by 7 publications

(5 citation statements)

References 36 publications

(32 reference statements)

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“…A4 and B4). Thus, these results demonstrated a divalent cation-dependent differential partitioning of a pool of tubulin and the CK2 responsible for its phosphorylation, as previously reported by Uzcanga et al (2003). Equivalent results were attained throughout the entire epimastigote growth curve (4-16 days).…”

Section: Other Proceduressupporting

confidence: 87%

“…Divalent cations hindered the solubility of both parasite proteins, as previously reported by Uzcanga et al (2003), and they co-eluted using various chromatographic separations on the basis of parameters as different as charge (anion and cation exchange) and size (gel filtration). Not even phosphocellulose, which has been reported as a very useful resin to purify casein kinases from mammalian , and chymotrypsinogen A (Chy, 25 kDa, 20 .…”

Section: I S C U S I O Nmentioning

confidence: 83%

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Tight binding between a pool of the heterodimeric α/β tubulin and a protein kinase CK2 inTrypanosoma cruziepimastigotes

et al. 2005

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Tubulin is the predominant phosphoprotein in Trypanosoma cruzi epimastigotes and is phosphorylated by a protein kinase CK2. Interestingly, the presence or absence of divalent cations affected the solubilization of a pool of the parasite tubulin and the CK2 responsible for its phosphorylation. This fraction of tubulin and its kinase co-eluted using phosphocellulose, DEAE-Sepharose and Sephacryl S-300 chromatographies. Anti-alpha tubulin antibodies co-immunoprecipitated both tubulin and the CK2 responsible for its phosphorylation, and anti-CK2 alpha-subunit antibodies immunoprecipitated radioactively labelled alpha and beta tubulin from phosphorylated epimastigote homogenates. Additionally, native polyacrylamide gel electrophoresis of the purified and radioactively labelled fraction containing tubulin and its kinase demonstrated the phosphorylation of a unique band that reacted with both anti-CK2 alpha-subunit and anti-tubulin antibodies. Together, these results establish a strong interaction between a pool of the heterodimeric alpha/beta tubulin and a CK2 in this parasite. Hydrodynamic measurements indicated that the T. cruzi tubulin-CK2 complex is globular with an estimated size of 145.4-147.5 kDa.

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Section: Other Proceduressupporting

confidence: 87%

Section: I S C U S I O Nmentioning

confidence: 83%

Tight binding between a pool of the heterodimeric α/β tubulin and a protein kinase CK2 inTrypanosoma cruziepimastigotes

et al. 2005

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“…In addition, the parasite CK2 was not purified to homogeneity, and inhibitors' IC 50 values may change according to the purity of the material. Consistently, tubulin has been reported to be the major phosphoprotein in another trypanosome, T. cruzi (Casas et al 2002), and likewise, T. cruzi tubulin has also been shown to be phosphorylated by a CK2-like activity (Casas et al 2002, Uzcanga et al 2003. In T. brucei, an evolutionarily more related parasite, two CK2 genes have been identified: CK2a1 and CK2a2 (Park et al 2002).…”

Section: Discussionmentioning

confidence: 60%

“…The specific activity of [γ-32 P] ATP was increased to 7200 cpm/pmol for this set of experiments. Protein Kinases in T. evansi • José Manuel Galán-Caridad et al 2002, Uzcanga et al 2003. It is plausible that the parasite CK2 also acts as a MAP regulating microtubule dynamics in trypanosomes through its direct physical association with tubulin, similar to what has been shown in higher eukaryotes.…”

Section: Discussionmentioning

confidence: 78%

Identification of casein kinase 1, casein kinase 2, and cAMP-dependent protein kinase-like activities in Trypanosoma evansi

Galán-Caridad

Calabokis

Uzcanga

et al. 2004

Mem. Inst. Oswaldo Cruz

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“…Insects can be plant feeders (Lustosa et al, 1999), predators (Zanuncio et al, 2003), or parasitoids (Matos et al 2004a,b) and they can transmit diseases (Contreras et al, 2002, Uzcanga et al, 2003.…”

Section: Introductionmentioning

confidence: 99%

Fertility and life expectancy of the predator Supputius cincticeps (Heteroptera: Pentatomidae) exposed to sublethal doses of permethrin

Zanuncio¹,

Zanúncio²,

Serrão³

et al. 2005

Biol. Res.

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The stinkbug Supputius cincticeps (Stål) (Heteroptera: Pentatomidae) can be found in agricultural and forest ecosystems feeding primarily on larvae of Coleoptera and Lepidoptera, where it can be exposed to insecticide applications. This study therefore aimed to evaluate the reproductive potential of'S. cincticeps after exposition to sublethal doses of permethrin (5.74 x 10 -3 , 5.74 x 10 -2 , 5.74 x 10 -1 , 5.74 and 57.44 ppb) through the use of a fertility life table. The development cycle of this predator was determined in order to calculate its net reproductive rate (R 0 ), the infinitesimal (r m ) and finite (λ) rates of increase in addition to mean generation time (T). The net reproductive (18.31), infinitesimal (r m ) (0.050) and finite (λ) (1.051) rates of increase were higher, while generation time (57.93 days) was shorter for S. cincticeps exposed to 5.74x10 -1 ppb of permethrin than in the control. This indicates a higher rate of population increase of this predator when exposed to this permethrin dose.

show abstract

Divalent cation hinder the solubilization of a tubulin kinase activity from Trypanosoma cruzi epimastigotes

Cited by 7 publications

References 36 publications

Tight binding between a pool of the heterodimeric α/β tubulin and a protein kinase CK2 inTrypanosoma cruziepimastigotes

Tight binding between a pool of the heterodimeric α/β tubulin and a protein kinase CK2 inTrypanosoma cruziepimastigotes

Identification of casein kinase 1, casein kinase 2, and cAMP-dependent protein kinase-like activities in Trypanosoma evansi

Fertility and life expectancy of the predator Supputius cincticeps (Heteroptera: Pentatomidae) exposed to sublethal doses of permethrin

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