Preparation of Small‐Molecule Microarrays by <i>trans</i>‐Cyclooctene Tetrazine Ligation and Their Application in the High‐Throughput Screening of Protein–Protein Interaction Inhibitors of Bromodomains

Zhang, Chong‐Jing; Tan, Chelsea Y. J.; Ge, Jingyan; Na, Zhenkun; Chen, Grace Y. J.; Uttamchandani, Mahesh; Sun, Hongyan; Yao, Shao Q.

doi:10.1002/anie.201307803

Cited by 42 publications

(32 citation statements)

References 39 publications

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“…Spotting of trans ‐cyclooctene‐PEG 4 ‐NHS (TCO‐PEG 4 ‐NHS) onto selected areas on the poly‐ l ‐lysine slide made the resulting TCO‐modified surface ready for MSN immobilization. We chose Tz–TCO ligation for MSN‐on‐a‐chip fabrication because this extremely fast and efficient bioorthogonal reaction has previously been shown to afford excellent performance in other SMM applications ,…”

Section: Resultsmentioning

confidence: 99%

“…This was likely due to the three‐dimensional nature and high surface areas/loading capacity of MSNs: instead of “one‐compound‐per‐spot” in traditional SMMs, in which compound immobilization efficiency is limited by available functional groups both on the compound and on the slide surface, a “many‐compounds‐per‐spot” scenario had resulted. This shows that our MSN‐on‐a‐chip could offer better detection sensitivity than traditional SMMs . Time‐dependent compound release profiles with Cy5@MSN‐on‐a‐chip carried out in phosphate buffered saline (PBS, pH 7.4) at 37 °C upon incubation for different periods of time after different UV irradiation times (365 nm) showed that most of the Cy5 dye encapsulating the MSNs (≈80 %) could be released with a 45 min UV irradiation time followed by another 30–60 min of buffer incubation at various MSN‐spotting concentrations (Figure A, B); no on‐chip dye leakage was observed in areas not exposed to UV irradiation even after prolonged buffer incubation (16 h; non‐UV in Figure B).…”

Section: Resultsmentioning

confidence: 99%

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MSN‐on‐a‐Chip: Cell‐Based Screenings Made Possible on a Small‐Molecule Microarray of Native Natural Products

Peng

Liew

et al. 2018

ChemBioChem

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Standard small-molecule microarrays (SMMs) are not well-suited for cell-based screening assays. Of the few attempts made thus far to render SMMs cell-compatible, all encountered major limitations. Here we report the first mesoporous silica nanoparticle (MSN)-on-a-chip platform capable of allowing high-throughput cell-based screening to be conducted on SMMs. By making use of a glass surface on which hundreds of MSNs, each encapsulated with a different native natural product, were immobilized in spatially defined manner, followed by on-chip mammalian cell growth and on-demand compound release, high-content screening was successfully carried out with readily available phenotypic detection methods. By combining this new MSN-on-a-chip system with small interfering RNA technology for the first time, we discovered that (+)-usniacin possesses synergistic inhibitory properties similar to those of olaparib (an FDA-approved drug) in BRCA1-knockdown cancer cells.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

MSN‐on‐a‐Chip: Cell‐Based Screenings Made Possible on a Small‐Molecule Microarray of Native Natural Products

Peng

Liew

et al. 2018

ChemBioChem

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“…13 Beside protein modification the reaction has been used in a whole range of applications6e including DNA modification11b, 14 and microarray preparation 11d. 15…”

Section: Introductionmentioning

confidence: 99%

Terminal Alkenes as Versatile Chemical Reporter Groups for Metabolic Oligosaccharide Engineering

Späte¹,

Schart²,

Schöllkopf³

et al. 2014

Chemistry A European J

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The Diels-Alder reaction with inverse electron demand (DAinv reaction) of 1,2,4,5-tetrazines with electron rich or strained alkenes was proven to be a bioorthogonal ligation reaction that proceeds fast and with high yields. An important application of the DAinv reaction is metabolic oligosaccharide engineering (MOE) which allows the visualization of glycoconjugates in living cells. In this approach, a sugar derivative bearing a chemical reporter group is metabolically incorporated into cellular glycoconjugates and subsequently derivatized with a probe by means of a bioorthogonal ligation reaction. Here, we investigated a series of new mannosamine and glucosamine derivatives with carbamate-linked side chains of varying length terminated by alkene groups and their suitability for labeling cell-surface glycans. Kinetic investigations showed that the reactivity of the alkenes in DAinv reactions increases with growing chain length. When applied to MOE, one of the compounds, peracetylated N-butenyloxycarbonylmannosamine, was especially well suited for labeling cell-surface glycans. Obviously, the length of its side chain represents the optimal balance between incorporation efficiency and speed of the labeling reaction. Sialidase treatment of the cells before the bioorthogonal labeling reaction showed that this sugar derivative is attached to the glycans in form of the corresponding sialic acid derivative and not epimerized to another hexosamine derivative to a considerable extent.

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“…[18][19][20][21] We identified 25 fragments that were used for the synthesis of a 625-member focused library by growing the 25 fragments with 25 diverse synthons (See the Supporting Information). [22][23] As shown in Figure 1, the library was screened against PCAF, thereby leading to the identification of compounds 8 a 1 b and 19 a 1 b.…”

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confidence: 99%

Identification of Covalent Bromodomain Binders through DNA Display of Small Molecules

et al. 2015

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The regulation of transcriptional programs by epigenetic readers (bromodomains) has been linked to the development of several pathologies. Notably, it has been implicated in the regulation of cellular growth and evasion of apoptosis, in cancer as well as in inflammation. The discovery of small-molecule probes to dissect the role of bromodomains is thus important. We demonstrate that specific cysteine residues conserved across the bromodomains can be harnessed for covalent trapping. We report the discovery of two small molecules that form a covalent bond with cysteine residues conserved across the bromodomain family, analyze the subset of bromodomains that can be addressed through covalent binding, and show proteomic analyses enabled by the enrichment of bromodomains from native lysates.

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Preparation of Small‐Molecule Microarrays by trans‐Cyclooctene Tetrazine Ligation and Their Application in the High‐Throughput Screening of Protein–Protein Interaction Inhibitors of Bromodomains

Cited by 42 publications

References 39 publications

MSN‐on‐a‐Chip: Cell‐Based Screenings Made Possible on a Small‐Molecule Microarray of Native Natural Products

MSN‐on‐a‐Chip: Cell‐Based Screenings Made Possible on a Small‐Molecule Microarray of Native Natural Products

Terminal Alkenes as Versatile Chemical Reporter Groups for Metabolic Oligosaccharide Engineering

Identification of Covalent Bromodomain Binders through DNA Display of Small Molecules

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