References of "Manoharan, Ganesh Babu 50030438"
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See detailPDE6D Inhibitors with a New Design Principle Selectively Block K‑Ras Activity
Siddiqui, Farid A.; Alam, Catharina; Rosenqvist, Petja et al

in ACS Omega (2020), 5(1), 832-842

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See detailMedium-Throughput Detection of Hsp90/Cdc37 Protein–Protein Interaction Inhibitors Using a Split Renilla Luciferase-Based Assay
Siddiqui, Farid A.; Parkkola, Hanna; Manoharan, Ganesh Babu UL et al

in SLAS Discovery (2019)

The protein-folding chaperone Hsp90 enables the maturation and stability of various oncogenic signaling proteins and is thus pursued as a cancer drug target. Folding in particular of protein kinases is ... [more ▼]

The protein-folding chaperone Hsp90 enables the maturation and stability of various oncogenic signaling proteins and is thus pursued as a cancer drug target. Folding in particular of protein kinases is assisted by the co-chaperone Cdc37. Several inhibitors against the Hsp90 ATP-binding site have been developed. However, they displayed significant toxicity in clinical trials. By contrast, the natural product conglobatin A has an exceptionally low toxicity in mice. It targets the protein–protein interface (PPI) of Hsp90 and Cdc37, suggesting that interface inhibitors have an interesting drug development potential. In order to identify inhibitors of the Hsp90/Cdc37 PPI, we have established a mammalian cell lysate-based, medium-throughput amenable split Renilla luciferase assay. This assay employs N-terminal and C-terminal fragments of Renilla luciferase fused to full-length human Hsp90 and Cdc37, respectively. We expect that our assay will allow for the identification of novel Hsp90/Cdc37 interaction inhibitors. Such tool compounds will help to evaluate whether the toxicity profile of Hsp90/Cdc37 PPI inhibitors is in general more favorable than that of ATP-competitive Hsp90 inhibitors. Further development of such tool compounds may lead to new classes of Hsp90 inhibitors with applications in cancer and other diseases. [less ▲]

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See detailHigh-throughput amenable fluorescence-assays to screen for calmodulin-inhibitors
Manoharan, Ganesh Babu UL; Kopra, Kari; Eskonen, Ville et al

in Analytical Biochemistry (2019), 572

The KRAS gene is highly mutated in human cancers and the focus of current Ras drug development efforts. Recently the interface between the C-terminus of K-Ras and calmodulin (CaM) was proposed as a target ... [more ▼]

The KRAS gene is highly mutated in human cancers and the focus of current Ras drug development efforts. Recently the interface between the C-terminus of K-Ras and calmodulin (CaM) was proposed as a target site to block K-Ras driven cancer cell stemness. We therefore aimed at developing a high-throughput amenable screening assay to identify novel CaM-inhibitors as potential K-Ras stemness-signaling disruptors. A modulated time-resolved Förster resonance energy transfer (mTR-FRET)-assay was developed and benchmarked against an identically designed fluorescence anisotropy (FA)-assay. In both assays, two CaM-binding peptides were labeled with Eu(III)-chelate or fluorescein and used as single-label reporter probes that were displaced from CaM upon competitor binding. Thus, peptidic and small molecule competitors with nanomolar to micromolar affinities to CaM could be detected, including a peptide that was derived from the C-terminus of K-Ras. In order to detect CaM-residue specific covalent inhibitors, a cell lysate-based Förster resonance energy transfer (FRET)-assay was furthermore established. This assay enabled us to measure the slow, residue-specific, covalent inhibition by ophiobolin A in the presence of other endogenous proteins. In conclusion, we have developed a panel of fluorescence-assays that allows identification of conventional and covalent CaM-inhibitors as potential disruptors of K-Ras driven cancer cell stemness. [less ▲]

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See detailThiazole- and selenazole-comprising high-affinity inhibitors possess bright microsecond-scale photoluminescence in complex with protein kinase CK2.
Vahter, Jurgen; Viht, Kaido; Uri, Asko et al

in Bioorganic & medicinal chemistry (2018), 26(18), 5062-5068

A previously disclosed protein kinase (PK) CK2-selective inhibitor 4-(2-amino-1,3-thiazol-5-yl)benzoic acid (ATB) and its selenium-containing counterpart (ASB) revealed remarkable room temperature ... [more ▼]

A previously disclosed protein kinase (PK) CK2-selective inhibitor 4-(2-amino-1,3-thiazol-5-yl)benzoic acid (ATB) and its selenium-containing counterpart (ASB) revealed remarkable room temperature phosphorescence when bound to the ATP pocket of the protein kinase CK2. Conjugation of these fragments with a mimic of CK2 substrate peptide resulted in bisubstrate inhibitors with increased affinity towards the kinase. Attachment of the fluorescent acceptor dye 5-TAMRA to the conjugates led to significant enhancement of intensity of long-lifetime (microsecond-scale) photoluminescence of both sulfur- and selenium-containing compounds. The developed photoluminescent probes make possible selective determination of the concentration of CK2 in cell lysates and characterization of CK2 inhibitors by means of time-gated measurement of photoluminescence. [less ▲]

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See detailPhosphorylation of Notch1 by Pim kinases promotes oncogenic signaling in breast and prostate cancer cells
Santio, Niina M.; Landor, Sebastian K.-J.; Vahtera, Laura et al

in Oncotarget (2016), 7(28), 43220-43238

Tumorigenesis is a multistep process involving co-operation between several deregulated oncoproteins. In this study, we unravel previously unrecognized interactions and crosstalk between Pim kinases and ... [more ▼]

Tumorigenesis is a multistep process involving co-operation between several deregulated oncoproteins. In this study, we unravel previously unrecognized interactions and crosstalk between Pim kinases and the Notch signaling pathway, with implications for both breast and prostate cancer. We identify Notch1 and Notch3, but not Notch2, as novel Pim substrates and demonstrate that for Notch1, the serine residue 2152 is phosphorylated by all three Pim family kinases. This target site is located in the second nuclear localization sequence (NLS) of the Notch1 intracellular domain (N1ICD), and is shown to be important for both nuclear localization and transcriptional activity of N1ICD. Phosphorylation-dependent stimulation of Notch1 signaling promotes migration of prostate cancer cells, balances glucose metabolism in breast cancer cells, and supports in vivo growth of both types of cancer cells on chick embryo chorioallantoic membranes. Furthermore, Pim-induced growth of orthotopic prostate xenografts in mice is associated with enhanced nuclear Notch1 activity. Finally, simultaneous inhibition of Pim and Notch abrogates the cellular responses more efficiently than individual treatments, opening up new vistas for combinatorial cancer therapy. [less ▲]

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See detailCombining chemical and genetic approaches for development of responsive FRET-based sensor systems for protein kinases
Manoharan, Ganesh Babu UL; Enkvist, Erki; Uri, Asko

in Biophysical Chemistry (2016), 211

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See detailBifunctional Ligands for Inhibition of Tight-Binding Protein–Protein Interactions
Ivan, Taavi; Enkvist, Erki; Viira, Birgit et al

in Bioconjugate Chemistry (2016), 27(8), 1900-1910

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See detailFRET-based screening assay using small-molecule photoluminescent probes in lysate of cells overexpressing RFP-fused protein kinases
Manoharan, Ganesh Babu UL; Enkvist, Erki; Kasari, Marje et al

in Analytical Biochemistry (2015)

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See detailPIM kinase-responsive microsecond-lifetime photoluminescent probes based on selenium-containing heteroaromatic tricycle
Ekambaram, Ramesh; Manoharan, Ganesh Babu UL; Enkvist, Erki et al

in RSC Advances (2015)

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See detailBenzoselenadiazole-based responsive long-lifetime photoluminescent probes for protein kinases
Ekambaram, Ramesh; Enkvist, Erki; Manoharan, Ganesh Babu UL et al

in Chemical Communications (2014)

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See detailActive site analysis of cis-epoxysuccinate hydrolase from Nocardia tartaricans using homology modeling and site-directed mutagenesis
Vinayagam, Vasu; Jayaraman, Kumaresan; Manoharan, Ganesh Babu UL et al

in Applied Microbiology and Biotechnology (2012), 93(6), 23772386

Detailed reference viewed: 51 (2 UL)