A Small-Molecule Microarray Approach for the Identification of E2 Enzyme Inhibitors in Ubiquitin-Like Conjugation Pathways

Zlotkowski, Katherine; Hewitt, William M.; Sinniah, Ranuka; Tropea, Joseph E.; Needle, D.; Lountos, G.T.; Barchi, Joseph J.; Waugh, David S.; Schneekloth, Jr. John S.

doi:10.1177/2472555216683937

Cited by 22 publications

(14 citation statements)

References 16 publications

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“…For WaterLOGSY experiments, a reference 1D- 1 H and 1D WaterLOGSY spectrum of 100 μM N -methyl- l -valine and 100 μM compound was collected, followed by a separate sample containing 1 μM KRAS , 100 μM N -methyl-L-valine, and 100 μM compound. All 1D- 1 H NMR spectra were recorded using the ‘zgesgp’ excitation sculpting water suppression pulse sequence from Bruker, with 128 scans ( 43 ). All data were processed and visualized with MestReNova software (Version 8.1.2-11880).…”

Section: Methodsmentioning

confidence: 99%

Characterization of clinically used oral antiseptics as quadruplex-binding ligands

Calabrese

Zlotkowski

Alden

et al. 2018

Nucleic Acids Research

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Approaches to characterize the nucleic acid-binding properties of drugs and druglike small molecules are crucial to understanding the behavior of these compounds in cellular systems. Here, we use a Small Molecule Microarray (SMM) profiling approach to identify the preferential interaction between chlorhexidine, a widely used oral antiseptic, and the G-quadruplex (G4) structure in the KRAS oncogene promoter. The interaction of chlorhexidine and related drugs to the KRAS G4 is evaluated using multiple biophysical methods, including thermal melt, fluorescence titration and surface plasmon resonance (SPR) assays. Chlorhexidine has a specific low micromolar binding interaction with the G4, while related drugs have weaker and/or less specific interactions. Through NMR experiments and docking studies, we propose a plausible binding mode driven by both aromatic stacking and groove binding interactions. Additionally, cancer cell lines harbouring oncogenic mutations in the KRAS gene exhibit increased sensitivity to chlorhexidine. Treatment of breast cancer cells with chlorhexidine decreases KRAS protein levels, while a KRAS gene transiently expressed by a promoter lacking a G4 is not affected. This work confirms that known ligands bind broadly to G4 structures, while other drugs and druglike compounds can have more selective interactions that may be biologically relevant.

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Section: Methodsmentioning

confidence: 99%

Characterization of clinically used oral antiseptics as quadruplex-binding ligands

Calabrese

Zlotkowski

Alden

et al. 2018

Nucleic Acids Research

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“…425 In addition, small-molecule microarray-based screening has also been successfully used to identify the inhibitor of the SUMO E2 enzyme Ubc9, and a few small-molecule inhibitors have been found. 426 Targeting DUB activity As mentioned, ubiquitination is a dynamic and reversible process, and DUBs mediate the removal and processing of Ub or polyubiquitin chains from ubiquitinated proteins. 427 Many DUBs have been found to participate in various events during the cell cycle progression, genomic instability regulation and tumorigenesis processes.…”

Section: Targeting E3 Enzymesmentioning

confidence: 99%

The role of ubiquitination in tumorigenesis and targeted drug discovery

Deng

Meng

Chen

et al. 2020

Sig Transduct Target Ther

415

308

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Ubiquitination, an important type of protein posttranslational modification (PTM), plays a crucial role in controlling substrate degradation and subsequently mediates the "quantity" and "quality" of various proteins, serving to ensure cell homeostasis and guarantee life activities. The regulation of ubiquitination is multifaceted and works not only at the transcriptional and posttranslational levels (phosphorylation, acetylation, methylation, etc.) but also at the protein level (activators or repressors). When regulatory mechanisms are aberrant, the altered biological processes may subsequently induce serious human diseases, especially various types of cancer. In tumorigenesis, the altered biological processes involve tumor metabolism, the immunological tumor microenvironment (TME), cancer stem cell (CSC) stemness and so on. With regard to tumor metabolism, the ubiquitination of some key proteins such as RagA, mTOR, PTEN, AKT, c-Myc and P53 significantly regulates the activity of the mTORC1, AMPK and PTEN-AKT signaling pathways. In addition, ubiquitination in the TLR, RLR and STING-dependent signaling pathways also modulates the TME. Moreover, the ubiquitination of core stem cell regulator triplets (Nanog, Oct4 and Sox2) and members of the Wnt and Hippo-YAP signaling pathways participates in the maintenance of CSC stemness. Based on the altered components, including the proteasome, E3 ligases, E1, E2 and deubiquitinases (DUBs), many molecular targeted drugs have been developed to combat cancer. Among them, small molecule inhibitors targeting the proteasome, such as bortezomib, carfilzomib, oprozomib and ixazomib, have achieved tangible success. In addition, MLN7243 and MLN4924 (targeting the E1 enzyme), Leucettamol A and CC0651 (targeting the E2 enzyme), nutlin and MI-219 (targeting the E3 enzyme), and compounds G5 and F6 (targeting DUB activity) have also shown potential in preclinical cancer treatment. In this review, we summarize the latest progress in understanding the substrates for ubiquitination and their special functions in tumor metabolism regulation, TME modulation and CSC stemness maintenance. Moreover, potential therapeutic targets for cancer are reviewed, as are the therapeutic effects of targeted drugs.Signal Transduction and Targeted Therapy (2020) 5:11; https://doi.

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“…70,71 Recent work has focused on developing novel approaches that center on small-molecule microarrays to identify inhibitors of the notoriously difficult-to-drug E2 conjugases. 72 Such studies may ultimately lead to the discovery of novel/clinically relevant inhibitors of Ubc9.…”

Section: Translational Approaches Targeting Sumoylationmentioning

confidence: 99%

SUMOylation in brain ischemia: Patterns, targets, and translational implications

Bernstock

Yang

et al. 2017

J Cereb Blood Flow Metab

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Post-translational protein modification by small ubiquitin-like modifier (SUMO) regulates a myriad of homeostatic and stress responses. The SUMOylation pathway has been extensively studied in brain ischemia. Convincing evidence is now at hand to support the notion that a major increase in levels of SUMOylated proteins is capable of inducing tolerance to ischemic stress. Therefore, the SUMOylation pathway has emerged as a promising therapeutic target for neuroprotection in the face of brain ischemia. Despite this, it is prudent to acknowledge that there are many key questions still to be addressed in brain ischemia related to SUMOylation. Accordingly, herein, we provide a critical review of literature within the field to summarize current knowledge and in so doing highlight pertinent translational implications of the SUMOylation pathway in brain ischemia.

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A Small-Molecule Microarray Approach for the Identification of E2 Enzyme Inhibitors in Ubiquitin-Like Conjugation Pathways

Cited by 22 publications

References 16 publications

Characterization of clinically used oral antiseptics as quadruplex-binding ligands

Characterization of clinically used oral antiseptics as quadruplex-binding ligands

The role of ubiquitination in tumorigenesis and targeted drug discovery

SUMOylation in brain ischemia: Patterns, targets, and translational implications

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