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Unveiling the significance of His160 residue in CK2α-CX-4945 interaction with implications for innovative diabetes treatment
No full textreservedDiabetes mellitus is a chronic metabolic disorder characterized by disturbances in the body's sugar metabolism due to inadequate insulin production or utilization. Current therapeutic approaches focus on management rather than a cure necessitating the pursuit of novel treatments. Understanding the molecular mechanisms of the disease may aid innovative therapy development. The “Diabetic Kinome” encompasses protein kinases including DYRK1A, GSK3β, and CK2 responsible for regulating functions involved in diabetes. Targeted inhibition of these kinases shows potential as an alternative treatment, although this approach is still under investigation. Challenges arise due to the significant similarity in the kinase domains of diabetic kinome members, which requires meticulous design of highly specific inhibitors. Precision in this regard is crucial for minimizing off-target effects and promoting the advancement of effective anti-diabetic treatment. CK2, also known as Casein Kinase 2, is a tetrameric enzyme consisting of two catalytic subunits (CK2α and CK2α') and two regulatory subunits (CK2β). It is a key regulator of various cellular processes. Additionally, the recent studies have indicated an involvement of this particular kinase in the endocrine functions of pancreatic β-cells. Therefore, CK2 becomes one of the molecular targets in the treatment of diabetes. One of the most renowned CK2 inhibitors is CX-4945, a potent ATP-competitive inhibitor. However, this inhibitor also binds to other kinases within the diabetic kinome such as DYRK1A, making it challenging to attribute the observed effect to a specific kinase. Further investigation is essential to elucidate the intricate interaction mechanism between the inhibitor and the kinases. The analysis of the available CK2α-CX-4945 and DYRK1A-CX-4945 structures indicated a crucial role of the His160 residue in CK2α, in facilitating the unique inhibitor-kinase interaction.
The exact role of His160 was further examined in the presented thesis. Two mutants of His160 (Ala and Glu) were created using site-directed mutagenesis, expressed in E. coli, and subsequently purified. The thermal shift assay analysis confirmed that introduced changes did not affect the stability of the protein. Moreover, these alterations did not affect the enzymatic activity, and the analysed CK2α variants remained equally active, as demonstrated in the ADP-GLO assay. Substitution of His160 by Ala or Glu affected the kinase-CX-4945 interaction. The strongest inhibition was observed for the wild-type protein, while both mutants (H160A and H160E) displayed higher IC50 values. Then, crystallization trials were conducted, yielding crystals of three CK2α kinase variants (WT, H160A, and H160E) in complex with the CX-4945 inhibitor. This will enable the explanation of the observed differences in the interaction between CK2α variants and CX-4945 at the molecular level.
These results emphasize His160 residue role in CK2α-CX-4945 interaction, guiding the design of potent and selective inhibitors. These findings hold promise for design innovative ATP-competitive inhibitors tailored for CK2α, advancing potential therapies for diabetes treatment
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Private foundations--taxable expenditures (Sec. 4945) /
No full text" ... discusses in detail the restrictions placed upon the activities of private foundations under [section] 4945. Enacted by the Tax Reform Act of 1969, [section] 4945 imposes an excise tax upon "taxable expenditures" of a private foundation, defined by [section] 4945(d) as including five separate categories of expenditures.
CX-4945 inhibits TGF-β1-induced EMT.
No full text<p>(A) The chemical structure of CX-4945. (B) The effect of CX-4945 on the viability of A549 cells in the presence of TGF-β1. Briefly, cells wre treated with TGF-β1 (5 ng/ml) alone or in combination with CX-4945 for 72 h, and then cell viability was measured. **, <i>p</i> < 0.01 (versus the control). The effect of CX-4945 on the TGF-β1-induced expression of epithelial and mesenchymal markers was evaluated by Western blot analysis (C) and immunocytochemistry (D). Briefly, after 24 h serum starvation, A549 cells were treated with TGF-β1 (5 ng/ml) alone or in combination with CX-4945 in media containing 0.1% FBS for 72 h. All scale bars represent 200 µm.</p
ATP-competitive inhibition of Clks by CX-4945.
No full text<p>(A) Diagram of the chemical structure of CX-4945 (B) Competition of CX-4945 with ATP for inhibition of Clk2 (C) Structural model of the Clk2 in complex with CX-4945. CX-4945 was predicted to bind to the ATP-binding site of Clk2. (D) The predicted binding mode between CX-4945 and Clk2. Amino acids in the active site are denoted by yellow sticks, and CX-4945 is indicated by green sticks. Hydrogen bonds are denoted as dotted black lines, and oxygen and nitrogen atoms are indicated as red and blue sticks, respectively.</p
Effect of CX-4945 on pre-mRNA splicing.
No full text<p>(A) CX-4945 induces expression of an unexpected CK2 α′ transcript, CK2 α′△. RT-PCR corresponding to exons 5–6–7 was performed with total RNA from DMSO- or CX-4945-treated 293T cells. (B) CX-4945 rapidly induces the expression of CK2 α′△. 293T cells were treated with CX-4945 for the times indicated prior to RNA preparation and RT-PCR analysis. GAPDH mRNA was analyzed as a control. (C) Analysis of the DNA sequence of CK2 α′△ revealed that it represents the exon6-skipped product (CK2 α′△exon6) of CK2 α′ pre-mRNA. The normally spliced (CK2 α′) and alternatively spliced (CK2 α′△exon6) products are illustrated schematically.</p
Cinemática de NGC 4945
No full textSe encuentra en elaboración un análisis interferométrico del campo de velocidades de NGC 4945 basado en más de 150 velocidades radiales repartidas en su cuerpo principal y determinadas de varios interferogramas Hα obtenidos en Bosque Alegre.Asociación Argentina de Astronomí
Drug combination using dasatinib and CX-4945.
No full text<p><b>A.</b> The Chou-Talalay method [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144126#pone.0144126.ref078" target="_blank">78</a>] was used to perform drug combination studies of dasatinib and CX-4945. The points represent the average viability ± standard error of mean following 72 h of drug treatment at the indicated concentrations of CX-4945 (•) and CX-4945 + dasatinib (◆; constant molar ratio of 20:1 of CX-4945:dasatinib) for the various EOC cell lines as a percentage of vehicle treated cells. The curve-fit lines were generated using non-linear regression analysis in GraphPad Prism. Data for the other molar ratios that were evaluated are presented in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144126#pone.0144126.s002" target="_blank">S2 Fig</a>. B.</b> The dose response data were used to calculate the Combination Index (CI) values for each cell line at the various molar ratios using CalcuSyn software [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144126#pone.0144126.ref079" target="_blank">79</a>]. CI values less than 1 suggest that the drugs are working synergistically. Shown is the average calculated CI value ± standard error of the mean.</p
Condiciones físicas en NGC 4945
Get PDFMediante observaciones espectroscópicas se han estudiado las variaciones de las condiciones físicas del gas ionizado en NGC 4945, encontrándose variaciones radiales de densidad y temperatura semejantes a las encontradas en otras galaxias de tipo semejante.Asociación Argentina de Astronomí
Far-infrared observations of NGC 4945 and the Circinus galaxy
No full textThe study reports observations of NGC 4945 and the Circinus galaxy carried out with a 1-m balloon-borne FIR telescope. Pointed observations of IRAS, using survey detectors, were deconvolved to get higher angular resolution intensity maps at 12, 25, 60, and 100 microns, respectively, for both galaxies. In NGC 4945, the nucleus was found to be cooler than its immediate surroundings. Whereas both galaxies show a prominent central source, only NGC 4945 shows extended disk emission in all bands. The central source in both galaxies was found to be somewhat more extended than a point source at wavelengths 50 microns or greater, the resulting deconvolved size being about 40 arcsec. The 150-micron upper limit for the Circinus galaxy implies a steeper wavelength dependence of dust emissivity than is the case for NGC 4945
CX-4945 inhibits TGF-β1-induced Wnt signaling.
No full text<p>The effect of CX-4945 on TGF-β1-induced Wnt activation was evaluated by measuring the transcriptional activity of β-catenin (A) and its expression in cytosolic and nuclear fractions (B). For Western blot analysis, A549 cells were treated with TGF-β1 (5 ng/ml) alone or in combination with CX-4945 in media containing 0.1% FBS for 48 h. Actin was used as a loading control. (C) Nuclear translocation of β-catenin and its inhibition by CX-4945 were confirmed by immunocytochemistry. Nuclei were counterstained with Hoechst 33342. All scale bars represent 20 µm.</p
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