Functional display of heterotetrameric human protein kinase CK2 on Escherichia coli: a novel tool for drug discovery

Background: Human protein kinase CK2 represents a novel therapeutic target for neoplastic diseases. Inhibitors are in need to explore the druggability and the therapeutic options of this enzyme. A bottleneck in the search for new inhibitors is the availability of the target for testing. Therefore an assay was developed to provide easy access to CK2 for discovery of novel inhibitors. Results: Autodisplay was used to present human CK2 on the surface of Escherichia coli. Heterotetrameric CK2 consists of two subunits, α and β, which were displayed individually on the surface. Co-display of CK2α and CK2β on the cell surface led to the formation of functional holoenzyme, as demonstrated by NaCl dependency of enzymatic activity, which differs from that of the catalytic subunit CK2α without β. In addition interaction of CK2α and CK2β at the cell surface was confirmed by co-immunoprecipitation assays. Surface displayed CK2 holoenzyme enabled an easy IC50 value determination. The IC50 values for the known CK2 inhibitors TBB and Silmitasertib were determined to be 50 and 3.3 nM, respectively. Conclusion: Surface-displayed CK2α and CK2β assembled on the cell surface of E. coli to an active tetrameric holoenzyme. The whole-cell CK2 autodisplay assay as developed is suitable for inhibition studies. Furthermore, it can be used to determine quantitative CK2 inhibition data such as IC50 values. In summary, this is the first report on the functional surface display of a heterotetrameric enzyme on E. coli.<br

The DEEP2 Galaxy Redshift Survey: Environments of Poststarburst Galaxies at z~0.1 and z~0.8

Postststarburst (K+A) galaxies are candidates for galaxies in transition from a star-forming phase to a passively-evolving phase. We have spectroscopically identified large samples of K+A galaxies both in the SDSS at z~0.1 and in the DEEP2 survey at z~0.8, using a robust selection method based on a cut in Hbeta emission rather than the more problematic [OII] 3727. Based on measurements of the overdensity of galaxies around each object, we find that K+A galaxies brighter than 0.4L*_B at low-z have a similar, statistically indistinguishable environment distribution as blue galaxies, preferring underdense environments, but dramatically different from that of red galaxies. However, at higher-z, the environment distribution of K+A galaxies is more similar to red galaxies than to blue galaxies. We conclude that the quenching of star formation and the build-up of the red sequence through the K+A phase is happening in relatively overdense environments at z~1 but in relatively underdense environments at z~0. Although the relative environments where quenching occurs are decreasing with time, the corresponding absolute environment may have stayed the same along with the quenching mechanisms, because the mean absolute environments of all galaxies has to grow with time. In addition, we do not find any significant dependence on luminosity in the environment distribution of K+As. The existence of a large K+A population in the field at both redshifts indicates that cluster-specific mechanisms cannot be the dominant route by which these galaxies are formed. We also demonstrates that studying K+A-environment relations by measuring the K+A fraction in different environments is highly non-robust. Statistical comparisons of the overall environment distributions of different populations are much better behaved.Comment: 21 pages, 13 figures, 4 tables, submitted to MNRAS; v2: major revision in Sec 5, 6, 7, and 9. Implemented robust statistical techniques in place of K+A fraction measurements; conclusions on the environment distribution of poststarbursts at z~0.8 have changed. Completely new discussion added. v3: minor changes matching the accepted versio

Synthesis andIn Vitro Evaluation of 3-(1-Azolylmethyl)-1H-indoles and 3-(1-Azolyl-1-phenylmethyl)-1H-indoles as Inhibitors of P450 arom

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Diacritic Binding of an Indenoindole Inhibitor by CK2 alpha Paralogs Explored by a Reliable Path to Atomic Resolution CK2 alpha ' Structures

CK2 alpha and CK2 alpha' are the two isoforms of the catalytic subunit of human protein kinase CK2, an important target for cancer therapy. They have similar, albeit not identical functional and structural properties, and were occasionally reported to be inhibited with distinct efficacies by certain ATP-competitive ligands. Here, we present THN27, an indeno[1,2-b] indole derivative, as a further inhibitor with basal isoform selectivity. The selectivity disappears when measured using CK2 alpha/CK2 alpha' complexes with CK2 beta, the regulatory CK2 subunit. Co-crystal structures of THN27 with CK2 alpha and CK2 alpha' reveal that subtle differences in the conformational variability of the inter-domain hinge region are correlated with the observed effect. In the case of CK2 alpha', a crystallographically problematic protein so far, this comparative structural analysis required the development of an experimental strategy that finally enables atomic resolution structure determinations with ab initio phasing of potentially any ATP-competitive CK2 inhibitor and possibly many non-ATP-competitive ligands as well bound to CK2 alpha'

QSAR Model of Indeno[1,2-b]indole Derivatives and Identification of N-isopentyl-2-methyl-4,9-dioxo-4,9-Dihydronaphtho[2,3-b]furan-3-carboxamide as a Potent CK2 Inhibitor

Casein kinase II (CK2) is an intensively studied enzyme, involved in different diseases, cancer in particular. Different scaffolds were used to develop inhibitors of this enzyme. Here, we report on the synthesis and biological evaluation of twenty phenolic, ketonic, and para-quinonic indeno[1,2-b]indole derivatives as CK2 inhibitors. The most active compounds were 5-isopropyl-1-methyl-5,6,7,8-tetrahydroindeno[1,2-b]indole-9,10-dione 4h and 1,3-dibromo-5-isopropyl-5,6,7,8-tetrahydroindeno[1,2-b]indole-9,10-dione 4w with identical IC50 values of 0.11 µM. Furthermore, the development of a QSAR model based on the structure of indeno[1,2-b]indoles was performed. This model was used to predict the activity of 25 compounds with naphtho[2,3-b]furan-4,9-dione derivatives, which were previously predicted as CK2 inhibitors via a molecular modeling approach. The activities of four naphtho[2,3-b]furan-4,9-dione derivatives were determined in vitro and one of them (N-isopentyl-2-methyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-3-carboxamide) turned out to inhibit CK2 with an IC50 value of 2.33 µM. All four candidates were able to reduce the cell viability by more than 60% after 24 h of incubation using 10 µM