Structured Review

BPS Bioscience fluorogenic hdac1 assay kit
Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 <t>(HDAC1)</t> enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p
Fluorogenic Hdac1 Assay Kit, supplied by BPS Bioscience, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fluorogenic hdac1 assay kit/product/BPS Bioscience
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
fluorogenic hdac1 assay kit - by Bioz Stars, 2021-07
86/100 stars

Images

1) Product Images from "A New Histone Deacetylase Inhibitor, MHY219, Inhibits the Migration of Human Prostate Cancer Cells via HDAC1"

Article Title: A New Histone Deacetylase Inhibitor, MHY219, Inhibits the Migration of Human Prostate Cancer Cells via HDAC1

Journal: Biomolecules & Therapeutics

doi: 10.4062/biomolther.2015.026

Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 (HDAC1) enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p
Figure Legend Snippet: Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 (HDAC1) enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p

Techniques Used: Histone Deacetylase Assay, Activity Assay, Expressing, HDAC Activity Assay

2) Product Images from "Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex"

Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

Journal: eLife

doi: 10.7554/eLife.37231

Nucleosomes with hydroxamic acid Lys analogs were potent competitive inhibitors of LHC-catalyzed deacetylation of Kac peptide substrate. ( A ) A model for nucleosomes containing hydroxamic acid acetyl-Lys analogs (KHd) on the H3 tail and coordinating to the active site Zn 2+ of HDAC1 in LHC. ( B ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified nucleosomes. ( C ) Kac and unacetylated nucleosomes (up to 400 nM) do not inhibit LHC catalyzed peptide deacetylation. ( D ) Dixon plot of LHC inhibition by H3K14Hd-modified nucleosomes shows competitive inhibition versus peptide deacetylation with K i = 35 nM. ( E ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified semisynthetic histone H3s. ( F ) Summary of the K i values calculated from IC 50 values, (n = 3 for all measurements); values shown are ± S.E.M.
Figure Legend Snippet: Nucleosomes with hydroxamic acid Lys analogs were potent competitive inhibitors of LHC-catalyzed deacetylation of Kac peptide substrate. ( A ) A model for nucleosomes containing hydroxamic acid acetyl-Lys analogs (KHd) on the H3 tail and coordinating to the active site Zn 2+ of HDAC1 in LHC. ( B ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified nucleosomes. ( C ) Kac and unacetylated nucleosomes (up to 400 nM) do not inhibit LHC catalyzed peptide deacetylation. ( D ) Dixon plot of LHC inhibition by H3K14Hd-modified nucleosomes shows competitive inhibition versus peptide deacetylation with K i = 35 nM. ( E ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified semisynthetic histone H3s. ( F ) Summary of the K i values calculated from IC 50 values, (n = 3 for all measurements); values shown are ± S.E.M.

Techniques Used: Inhibition, Modification

The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.
Figure Legend Snippet: The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

Techniques Used: Sequencing

3) Product Images from "Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors"

Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors

Journal: Nature Communications

doi: 10.1038/s41467-017-02242-4

Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments
Figure Legend Snippet: Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments

Techniques Used: Activity Assay, Staining, Purification, Size-exclusion Chromatography, Produced, Inhibition, Generated, Mass Spectrometry, Western Blot

4) Product Images from "Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex"

Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

Journal: eLife

doi: 10.7554/eLife.37231

LHC and isolated HDAC1 deacetylation of nucleosomes and isolated histone H3 containing Kac. ( A ) His-FLAG-HDAC1 (50 nM) showed no detectable deacetylation of H3K9ac nucleosome substrate (100 nM). ( B ) His-FLAG-HDAC1 (5 nM) showed robust deacetylation of both H3K9ac and H3K14ac histone H3 substrates (1 μM). ( C ) Deacetylation of the H3K9K14K18 triacetylated histone H3 (1 μM) by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes (lacking H3K4me2 modification) by 20 nM LHC. Each of these panels shows western blots with the relevant site-specific and total H3 Abs. (n = 3 for all measurements except n = 2 for A and B).
Figure Legend Snippet: LHC and isolated HDAC1 deacetylation of nucleosomes and isolated histone H3 containing Kac. ( A ) His-FLAG-HDAC1 (50 nM) showed no detectable deacetylation of H3K9ac nucleosome substrate (100 nM). ( B ) His-FLAG-HDAC1 (5 nM) showed robust deacetylation of both H3K9ac and H3K14ac histone H3 substrates (1 μM). ( C ) Deacetylation of the H3K9K14K18 triacetylated histone H3 (1 μM) by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes (lacking H3K4me2 modification) by 20 nM LHC. Each of these panels shows western blots with the relevant site-specific and total H3 Abs. (n = 3 for all measurements except n = 2 for A and B).

Techniques Used: Isolation, Modification, Western Blot

Demethylation kinetics of H3K4me2 nucleosomes (100 nM) by LHC (400 nM) quantified by western blots. ( A ) The H3K4me2 nucleosome with 185 bp dsDNA was slowly demethylated by the CoREST complex while nucleosome demethylation with with 146 bp dsDNA was below the level of detection. ( B ) The LSD1 and HDAC1 dual inhibitor Corin (20 μM) inhibited the demethylation of the 185 bp nucleosome substrate; (n = 3 for A and n = 2 for B); error bars represent S.E.M.
Figure Legend Snippet: Demethylation kinetics of H3K4me2 nucleosomes (100 nM) by LHC (400 nM) quantified by western blots. ( A ) The H3K4me2 nucleosome with 185 bp dsDNA was slowly demethylated by the CoREST complex while nucleosome demethylation with with 146 bp dsDNA was below the level of detection. ( B ) The LSD1 and HDAC1 dual inhibitor Corin (20 μM) inhibited the demethylation of the 185 bp nucleosome substrate; (n = 3 for A and n = 2 for B); error bars represent S.E.M.

Techniques Used: Western Blot

The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.
Figure Legend Snippet: The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

Techniques Used: Sequencing

Quantification and curve fitting of the deacetylation assays. ( A ) Deacetylation of 1 μM Kac histone H3 by 1 nM LHC. ( B ) Deacetylation of 100 nM H3K9K14K18 triacetylated nucleosomes by 40 nM LHC. ( C ) Deacetylation of 1 μM H3K9K14K18 triacetylated histone H3 by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes without H3K4me2 by 20 nM LHC. V/[E]=0.14 ± 0.0045 min −1 . ( E ) The deacetylation of 1 μM Kac histone H3 by 5 nM His-FLAG-HDAC1; (n = 3 for all measurements except n = 2 for A and C); Error bars represent S.E.M.
Figure Legend Snippet: Quantification and curve fitting of the deacetylation assays. ( A ) Deacetylation of 1 μM Kac histone H3 by 1 nM LHC. ( B ) Deacetylation of 100 nM H3K9K14K18 triacetylated nucleosomes by 40 nM LHC. ( C ) Deacetylation of 1 μM H3K9K14K18 triacetylated histone H3 by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes without H3K4me2 by 20 nM LHC. V/[E]=0.14 ± 0.0045 min −1 . ( E ) The deacetylation of 1 μM Kac histone H3 by 5 nM His-FLAG-HDAC1; (n = 3 for all measurements except n = 2 for A and C); Error bars represent S.E.M.

Techniques Used:

5) Product Images from "Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors"

Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors

Journal: Nature Communications

doi: 10.1038/s41467-017-02242-4

Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments
Figure Legend Snippet: Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments

Techniques Used: Activity Assay, Staining, Purification, Size-exclusion Chromatography, Produced, Inhibition, Generated, Mass Spectrometry, Western Blot

Related Articles

other:

Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex
Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

Inhibition:

Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors
Article Snippet: .. HDAC1 inhibition assay HDAC1 inhibitory activity was determined for each compound using the Fluorogenic HDAC1 assay kit available from BPS Bioscience (50061). ..

Activity Assay:

Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors
Article Snippet: .. HDAC1 inhibition assay HDAC1 inhibitory activity was determined for each compound using the Fluorogenic HDAC1 assay kit available from BPS Bioscience (50061). ..

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension
Article Snippet: Mice were sacrificed at the end of the study by CO2 inhalation. .. Fluorogenic class I HDAC enzyme activities To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols. ..

Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors
Article Snippet: .. HDAC1 inhibitory activity was determined for each compound using the Fluorogenic HDAC1 assay kit available from BPS Bioscience (50061). ..

Article Title: Preclinical anti-arthritic study and pharmacokinetic properties of a potent histone deacetylase inhibitor MPT0G009
Article Snippet: The labeled secondary antibodies were horseradish peroxidase- or FITC-conjugated anti-mouse or anti-rabbit IgG antibodies (Jackson ImmunoResearch Inc., West Grove, PA, USA). .. Fluorogenic HDAC1, 2, 3, 4, 6 or 8 assay kits were from BPS Bioscience Corp. (San Diego, CA, USA), colorimetric HDAC activity kits from BioVision Inc. (Milpitas, CA, USA), PGE2 immunoassay kits from R & D Systems (Minneapolis, MN, USA) and IL-6 ELISA kits from eBioscience Inc. (San Diego, CA, USA). .. The pGL4.32[luc2P/ NF-κ B-RE/ Hygro] and pGL4.30[luc2P/ NFAT-RE/ Hygro] plasmids were from Promega Corp. (Madison, HI, USA) and the HDAC1-Flag (plasmid 13820) and HDAC6-Flag (plasmid 13823) plasmids from Addgene Inc. (Cambridge, MA, USA).

Article Title: A New Histone Deacetylase Inhibitor, MHY219, Inhibits the Migration of Human Prostate Cancer Cells via HDAC1
Article Snippet: .. HDAC1 activity assay HDAC1 enzyme activity was assessed using a fluorogenic HDAC1 assay kit purchased from BPS Bioscience (San Diego, CA, USA) according to the manufacturer’s instructions. .. Briefly, HDAC1 enzyme was incubated with vehicle or various concentrations of TsA, SAHA, or MHY219 at 37°C for 30 min in the presence of an HDAC1 fluorometric substrate.

Mass Spectrometry:

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension
Article Snippet: Mice were sacrificed at the end of the study by CO2 inhalation. .. Fluorogenic class I HDAC enzyme activities To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols. ..

Enzymatic Assay:

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension
Article Snippet: Mice were sacrificed at the end of the study by CO2 inhalation. .. Fluorogenic class I HDAC enzyme activities To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols. ..

Enzyme-linked Immunosorbent Assay:

Article Title: Preclinical anti-arthritic study and pharmacokinetic properties of a potent histone deacetylase inhibitor MPT0G009
Article Snippet: The labeled secondary antibodies were horseradish peroxidase- or FITC-conjugated anti-mouse or anti-rabbit IgG antibodies (Jackson ImmunoResearch Inc., West Grove, PA, USA). .. Fluorogenic HDAC1, 2, 3, 4, 6 or 8 assay kits were from BPS Bioscience Corp. (San Diego, CA, USA), colorimetric HDAC activity kits from BioVision Inc. (Milpitas, CA, USA), PGE2 immunoassay kits from R & D Systems (Minneapolis, MN, USA) and IL-6 ELISA kits from eBioscience Inc. (San Diego, CA, USA). .. The pGL4.32[luc2P/ NF-κ B-RE/ Hygro] and pGL4.30[luc2P/ NFAT-RE/ Hygro] plasmids were from Promega Corp. (Madison, HI, USA) and the HDAC1-Flag (plasmid 13820) and HDAC6-Flag (plasmid 13823) plasmids from Addgene Inc. (Cambridge, MA, USA).

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    BPS Bioscience fluorogenic hdac1 assay kit
    Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 <t>(HDAC1)</t> enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p
    Fluorogenic Hdac1 Assay Kit, supplied by BPS Bioscience, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluorogenic hdac1 assay kit/product/BPS Bioscience
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fluorogenic hdac1 assay kit - by Bioz Stars, 2021-07
    86/100 stars
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    Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 (HDAC1) enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p

    Journal: Biomolecules & Therapeutics

    Article Title: A New Histone Deacetylase Inhibitor, MHY219, Inhibits the Migration of Human Prostate Cancer Cells via HDAC1

    doi: 10.4062/biomolther.2015.026

    Figure Lengend Snippet: Effect of MHY219 and SAHA on histone deacetylase (HDAC) 1 activity and expression levels of HDACs in prostate cancer cells. (A) Histone deacetylase 1 (HDAC1) enzyme activity was measured by using a fluorometric HDAC activity assay kit. This result represents the percentage of activity compared to control cells in each group, respectively. Results are expressed as mean ± standard error of the mean (S.E.M.) of three independent experiments. * p

    Article Snippet: HDAC1 activity assay HDAC1 enzyme activity was assessed using a fluorogenic HDAC1 assay kit purchased from BPS Bioscience (San Diego, CA, USA) according to the manufacturer’s instructions.

    Techniques: Histone Deacetylase Assay, Activity Assay, Expressing, HDAC Activity Assay

    Nucleosomes with hydroxamic acid Lys analogs were potent competitive inhibitors of LHC-catalyzed deacetylation of Kac peptide substrate. ( A ) A model for nucleosomes containing hydroxamic acid acetyl-Lys analogs (KHd) on the H3 tail and coordinating to the active site Zn 2+ of HDAC1 in LHC. ( B ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified nucleosomes. ( C ) Kac and unacetylated nucleosomes (up to 400 nM) do not inhibit LHC catalyzed peptide deacetylation. ( D ) Dixon plot of LHC inhibition by H3K14Hd-modified nucleosomes shows competitive inhibition versus peptide deacetylation with K i = 35 nM. ( E ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified semisynthetic histone H3s. ( F ) Summary of the K i values calculated from IC 50 values, (n = 3 for all measurements); values shown are ± S.E.M.

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: Nucleosomes with hydroxamic acid Lys analogs were potent competitive inhibitors of LHC-catalyzed deacetylation of Kac peptide substrate. ( A ) A model for nucleosomes containing hydroxamic acid acetyl-Lys analogs (KHd) on the H3 tail and coordinating to the active site Zn 2+ of HDAC1 in LHC. ( B ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified nucleosomes. ( C ) Kac and unacetylated nucleosomes (up to 400 nM) do not inhibit LHC catalyzed peptide deacetylation. ( D ) Dixon plot of LHC inhibition by H3K14Hd-modified nucleosomes shows competitive inhibition versus peptide deacetylation with K i = 35 nM. ( E ) Dose-dependent inhibition of LHC catalyzed peptide deacetylation by KHd-modified semisynthetic histone H3s. ( F ) Summary of the K i values calculated from IC 50 values, (n = 3 for all measurements); values shown are ± S.E.M.

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: Inhibition, Modification

    The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: Sequencing

    Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments

    Journal: Nature Communications

    Article Title: Targeting the CoREST complex with dual histone deacetylase and demethylase inhibitors

    doi: 10.1038/s41467-017-02242-4

    Figure Lengend Snippet: Dual inhibitors exhibit unique activity against the CoREST complex. a Coomassie stained gel depicting the three components of the CoREST ternary complex after purification by size exclusion chromatography. b Dose response produced by inhibition of LSD1 as part of the CoREST complex by corin and structurally matched compound 7 . c Inhibition curves generated for corin and MS-275 against HDAC1 as part of the CoREST complex. d Corin inhibited the deacetylation of reconstituted nucleosomes by the CoREST complex as determined by Western blot (CoREST complex = 100 nM, nucleosome = 100 nM). Data (mean ± SEM) are representative of at least two independent experiments

    Article Snippet: HDAC1 inhibitory activity was determined for each compound using the Fluorogenic HDAC1 assay kit available from BPS Bioscience (50061).

    Techniques: Activity Assay, Staining, Purification, Size-exclusion Chromatography, Produced, Inhibition, Generated, Mass Spectrometry, Western Blot

    LHC and isolated HDAC1 deacetylation of nucleosomes and isolated histone H3 containing Kac. ( A ) His-FLAG-HDAC1 (50 nM) showed no detectable deacetylation of H3K9ac nucleosome substrate (100 nM). ( B ) His-FLAG-HDAC1 (5 nM) showed robust deacetylation of both H3K9ac and H3K14ac histone H3 substrates (1 μM). ( C ) Deacetylation of the H3K9K14K18 triacetylated histone H3 (1 μM) by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes (lacking H3K4me2 modification) by 20 nM LHC. Each of these panels shows western blots with the relevant site-specific and total H3 Abs. (n = 3 for all measurements except n = 2 for A and B).

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: LHC and isolated HDAC1 deacetylation of nucleosomes and isolated histone H3 containing Kac. ( A ) His-FLAG-HDAC1 (50 nM) showed no detectable deacetylation of H3K9ac nucleosome substrate (100 nM). ( B ) His-FLAG-HDAC1 (5 nM) showed robust deacetylation of both H3K9ac and H3K14ac histone H3 substrates (1 μM). ( C ) Deacetylation of the H3K9K14K18 triacetylated histone H3 (1 μM) by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes (lacking H3K4me2 modification) by 20 nM LHC. Each of these panels shows western blots with the relevant site-specific and total H3 Abs. (n = 3 for all measurements except n = 2 for A and B).

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: Isolation, Modification, Western Blot

    Demethylation kinetics of H3K4me2 nucleosomes (100 nM) by LHC (400 nM) quantified by western blots. ( A ) The H3K4me2 nucleosome with 185 bp dsDNA was slowly demethylated by the CoREST complex while nucleosome demethylation with with 146 bp dsDNA was below the level of detection. ( B ) The LSD1 and HDAC1 dual inhibitor Corin (20 μM) inhibited the demethylation of the 185 bp nucleosome substrate; (n = 3 for A and n = 2 for B); error bars represent S.E.M.

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: Demethylation kinetics of H3K4me2 nucleosomes (100 nM) by LHC (400 nM) quantified by western blots. ( A ) The H3K4me2 nucleosome with 185 bp dsDNA was slowly demethylated by the CoREST complex while nucleosome demethylation with with 146 bp dsDNA was below the level of detection. ( B ) The LSD1 and HDAC1 dual inhibitor Corin (20 μM) inhibited the demethylation of the 185 bp nucleosome substrate; (n = 3 for A and n = 2 for B); error bars represent S.E.M.

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: Western Blot

    The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: The CoREST complex and its H3 substrate histone modifications. ( A ) The CoREST complex (LHC) represses gene transcription by erasing histone modifications on nucleosomes. LSD1 demethylates H3K4me2 and HDAC1 deacetylates multiple Kac sites on histones which are activating marks. ( B ) The sequence of the N-terminal histone H3 tail and the Kme2 and Kac modifications studied in this paper. The short recognition sequence of F40 sortase is labelled in green.

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: Sequencing

    Quantification and curve fitting of the deacetylation assays. ( A ) Deacetylation of 1 μM Kac histone H3 by 1 nM LHC. ( B ) Deacetylation of 100 nM H3K9K14K18 triacetylated nucleosomes by 40 nM LHC. ( C ) Deacetylation of 1 μM H3K9K14K18 triacetylated histone H3 by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes without H3K4me2 by 20 nM LHC. V/[E]=0.14 ± 0.0045 min −1 . ( E ) The deacetylation of 1 μM Kac histone H3 by 5 nM His-FLAG-HDAC1; (n = 3 for all measurements except n = 2 for A and C); Error bars represent S.E.M.

    Journal: eLife

    Article Title: Lysine-14 acetylation of histone H3 in chromatin confers resistance to the deacetylase and demethylase activities of an epigenetic silencing complex

    doi: 10.7554/eLife.37231

    Figure Lengend Snippet: Quantification and curve fitting of the deacetylation assays. ( A ) Deacetylation of 1 μM Kac histone H3 by 1 nM LHC. ( B ) Deacetylation of 100 nM H3K9K14K18 triacetylated nucleosomes by 40 nM LHC. ( C ) Deacetylation of 1 μM H3K9K14K18 triacetylated histone H3 by 1 nM LHC. ( D ) Deacetylation of 100 nM H3K9ac nucleosomes without H3K4me2 by 20 nM LHC. V/[E]=0.14 ± 0.0045 min −1 . ( E ) The deacetylation of 1 μM Kac histone H3 by 5 nM His-FLAG-HDAC1; (n = 3 for all measurements except n = 2 for A and C); Error bars represent S.E.M.

    Article Snippet: His-FLAG-HDAC1 and Fluorogenic HDAC1 assay kit was purchased from BPS Bioscience (San Diego, CA).

    Techniques: