UHPLC‐MS‐based HDAC Assay Applied to Bio‐guided Microfractionation of Fungal Extracts

Zwick, Vincent; Allard, Pierre‐Marie; Ory, L; Simões-Pires, Cláudia A.; Marcourt, Laurence; Gindro, Katia; Wolfender, Jean‐Luc; Cuendet, Muriel

doi:10.1002/pca.2652

Cited by 5 publications

(2 citation statements)

References 45 publications

(50 reference statements)

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“…As described in Table 2, it exhibited higher activities than the positive control (ascorbic acid), which corresponds to the result of a previous report [44]. Gentisyl alcohol has also been reported to have proangiogenic, caspase inhibition, histone deacetylases inhibition, antibacterial (against methicillin-resistant Staphylococcus aureus ), antifungal (against Colletotrichum gloeosporioides ), antileishmanial (against Leishmania donovani ), and cytotoxic (against human breast and colon cancer cell lines, MCF-7 and HT-29, respectively, and against sea urchin Strongylocentrotus intermedius ) activity [45,46,47,48,49,50,51,52,53]. This is the first report of the production of gentisyl alcohol from the genus Arthrinium .…”

Section: Resultsmentioning

confidence: 72%

Diversity and Ecology of Marine Algicolous Arthrinium Species as a Source of Bioactive Natural Products

et al. 2018

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In our previous study, all Arthrinium isolates from Sargassum sp. showed high bioactivities, but studies on marine Arthrinium spp. are insufficient. In this study, a phylogenetic analysis of 28 Arthrinium isolates from seaweeds and egg masses of Arctoscopus japonicus was conducted using internal transcribed spacers, nuclear large subunit rDNA, β-tubulin, and translation elongation factor region sequences, and their bioactivities were investigated. They were analyzed as 15 species, and 11 of them were found to be new species. Most of the extracts exhibited radical-scavenging activity, and some showed antifungal activities, tyrosinase inhibition, and quorum sensing inhibition. It was implied that marine algicolous Arthrinium spp. support the regulation of reactive oxygen species in symbiotic algae and protect against pathogens and bacterial biofilm formation. The antioxidant from Arthrinium sp. 10 KUC21332 was separated by bioassay-guided isolation and identified to be gentisyl alcohol, and the antioxidant of Arthrinium saccharicola KUC21221 was identical. These results demonstrate that many unexploited Arthrinium species still exist in marine environments and that they are a great source of bioactive compounds.

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Section: Resultsmentioning

confidence: 72%

Diversity and Ecology of Marine Algicolous Arthrinium Species as a Source of Bioactive Natural Products

et al. 2018

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“…Typical methods for detection of deacetylation include fluorophore-conjugated substrates, coupled detection of produced free lysine, coupled detection of produced acetate, and HPLC or mass spectrometric detection of the peptide product. 22–27 Ideal reaction conditions should permit detection with any of these approaches while ensuring consistent results that are not method-dependent.…”

Section: Introductionmentioning

confidence: 99%

Chelatable trace zinc causes low, irreproducible KDAC8 activity

Toro

Edenfield

Hylton

et al. 2018

Analytical Biochemistry

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Acetylation is an important regulatory mechanism in cells, and emphasis is being placed on identifying substrates and small molecule modulators of this post-translational modification. However, the reported in vitro activity of the lysine deacetylase KDAC8 is inconsistent across experimental setups, even with the same substrate, complicating progress in the field. We detected trace levels of zinc, a known inhibitor of KDAC8 when present in excess, even in high-quality buffer reagents, at concentrations that are sufficient to significantly inhibit the enzyme under common reaction conditions. We hypothesized that trace zinc in solution could account for the observed variability in KDAC8 activity. We demonstrate that addition of chelators, including BSA, EDTA, and citrate, and/or the use of a phosphate-based buffer instead of the more common tris-based buffer, eliminates the inhibition from low levels of zinc as well as the dependence of specific activity on enzyme concentration. This results in high KDAC8 activity that is consistent across buffer systems, even using low concentrations of enzyme. We report conditions that are suitable for several assays to increase both enzyme activity and reproducibility. Our results have significant implications for approaches used to identify substrates and small molecule modulators of KDAC8 and interpretation of existing data.

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Natural Agents-Mediated Targeting of Histone Deacetylases

Farooqı

Naqvi

Perk

et al. 2017

Arch. Immunol. Ther. Exp.

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In the past few years, basic and clinical scientists have witnessed landmark achievements in many research projects, such as those conducted by the US National Institutes of Health Roadmap Epigenomics Mapping Consortium, the International Human Epigenome Consortium, The Cancer Genome Atlas Network and the International Cancer Genome Consortium, which have provided near-complete resolution of epigenetic landscape in different diseases. Furthermore, genome sequencing of tumors has provided compelling evidence related to frequent existence of mutations in readers, erasers and writers of epigenome in different cancers. Histone acetylation is an intricate mechanism modulated by two opposing sets of enzymes and deeply studied as a key biological phenomenon in 1964 by Vincent Allfrey and colleagues. The research group suggested that this protein modification contributed substantially in transcriptional regulation. Subsequently, histone deacetylases (HDACs), histone acetyltransferases and acetyl-Lys-binding proteins were identified as transcriptional mediators, which further deepened our comprehension regarding biochemical modifications. Overwhelmingly increasing high-impact research is improving our understanding of this molecularly controlled mechanism; moreover, quantification and identification of lysine acetylation by mass spectrometry has added new layers of information. We partition this multi-component review into how both activity and expression of HDAC are targeted using natural agents. We also set spotlight on how oncogenic fusion proteins tactfully utilize HDAC-associated nano-machinery to modulate expression of different genes and how HDAC inhibitors regulate TRAIL-induced apoptosis in cancer cells. HDAC inhibitors have been reported to upregulate expression of TRAIL receptors and protect TRAIL from proteasomal degradation. Deeper understanding of HDAC biology will be useful for stratification and selection of patients who are responders, non-responders and poor-responders for HDACi therapy, and for the rational design of combination studies using HDACi.

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UHPLC‐MS‐based HDAC Assay Applied to Bio‐guided Microfractionation of Fungal Extracts

Abstract: The UHPLC-ESI-MS/MS-based method using specific HDAC isoforms is suitable to isolate selective HDAC inhibitors by bio-guided fractionation of fungal strains. Also, it decreases potential interferences with natural products compared to the fluorescence-based assay.

Cited by 5 publications

References 45 publications

Diversity and Ecology of Marine Algicolous Arthrinium Species as a Source of Bioactive Natural Products

Diversity and Ecology of Marine Algicolous Arthrinium Species as a Source of Bioactive Natural Products

Chelatable trace zinc causes low, irreproducible KDAC8 activity

Natural Agents-Mediated Targeting of Histone Deacetylases

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