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Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor

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Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor. / Kristensen, Line Hyltoft; Nielsen, Anders Laerke; Helgstrand, Charlotte; Lees, Michael; Cloos, Paul; Kastrup, Jette Sandholm; Helin, Kristian; Olsen, Lars; Gajhede, Michael.

In: F E B S Journal, Vol. 279, No. 11, 06.2012, p. 1905-1914.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Kristensen, LH, Nielsen, AL, Helgstrand, C, Lees, M, Cloos, P, Kastrup, JS, Helin, K, Olsen, L & Gajhede, M 2012, 'Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor', F E B S Journal, vol. 279, no. 11, pp. 1905-1914. https://doi.org/10.1111/j.1742-4658.2012.08567.x

APA

Kristensen, L. H., Nielsen, A. L., Helgstrand, C., Lees, M., Cloos, P., Kastrup, J. S., ... Gajhede, M. (2012). Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor. F E B S Journal, 279(11), 1905-1914. https://doi.org/10.1111/j.1742-4658.2012.08567.x

Vancouver

Kristensen LH, Nielsen AL, Helgstrand C, Lees M, Cloos P, Kastrup JS et al. Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor. F E B S Journal. 2012 Jun;279(11):1905-1914. https://doi.org/10.1111/j.1742-4658.2012.08567.x

Author

Kristensen, Line Hyltoft ; Nielsen, Anders Laerke ; Helgstrand, Charlotte ; Lees, Michael ; Cloos, Paul ; Kastrup, Jette Sandholm ; Helin, Kristian ; Olsen, Lars ; Gajhede, Michael. / Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor. In: F E B S Journal. 2012 ; Vol. 279, No. 11. pp. 1905-1914.

Bibtex

@article{9169de0bb5894e80901a726be45947f2,
title = "Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor",
abstract = "Dynamic methylations and demethylations of histone lysine residues are important for gene regulation and are facilitated by histone methyltransferases and histone demethylases (HDMs). KDM5B/Jarid1B/PLU1 is an H3K4me3/me2 specific lysine demethylase belonging to the family of JmjC domain containing lysine specific HDMs (JHDMs). Several studies have linked KDM5B to breast, prostate and skin cancer, highlighting its potential as a drug target. However, most inhibitor studies have focused on other JHDMs, and inhibitors for KDM5B remain to be explored. Here, we report the expression, purification and characterization of the catalytic core of recombinant KDM5B (residues 1-769, ccKDM5B). We show that ccKDM5B, recombinantly expressed in insect cells, demethylates H3K4me3 and H3K4me2 in vitro. The kinetic characterization showed that ccKDM5B has a K(m) (app) value of 0.5 µM for its tri-methylated substrate H3(1-15)K4me3, a considerably increased apparent substrate affinity than reported for related histone demethylases. Despite the presence of a PHD domain, the catalytic activity was not affected by additional methylation at the H3K9 position, suggesting that in vitro chromatin cross talk between H3K4 and H3K9 does not occur for ccKDM5B. Inhibition studies of ccKDM5B showed both in vitro and in cell inhibition of ccKDM5B by 2,4-pyridinedicarboxylic acid (2,4-PDCA) with a potency similar to that reported for the histone demethylase KDM4C. Structure-guided sequence alignment indicated that the binding mode of 2,4-PDCA is conserved between KDM4A/C and KDM5B.",
keywords = "The Faculty of Pharmaceutical Sciences, BRIC",
author = "Kristensen, {Line Hyltoft} and Nielsen, {Anders Laerke} and Charlotte Helgstrand and Michael Lees and Paul Cloos and Kastrup, {Jette Sandholm} and Kristian Helin and Lars Olsen and Michael Gajhede",
note = "Keywords: catalysis, expression, inhibition, KDM5B, purification",
year = "2012",
month = "6",
doi = "10.1111/j.1742-4658.2012.08567.x",
language = "English",
volume = "279",
pages = "1905--1914",
journal = "F E B S Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "11",

}

RIS

TY - JOUR

T1 - Studies of H3K4me3 demethylation by KDM5B/Jarid1B/PLU1 reveals strong substrate recognition in vitro and identifies 2,4-pyridine-dicarboxylic acid as an in vitro and in cell inhibitor

AU - Kristensen, Line Hyltoft

AU - Nielsen, Anders Laerke

AU - Helgstrand, Charlotte

AU - Lees, Michael

AU - Cloos, Paul

AU - Kastrup, Jette Sandholm

AU - Helin, Kristian

AU - Olsen, Lars

AU - Gajhede, Michael

N1 - Keywords: catalysis, expression, inhibition, KDM5B, purification

PY - 2012/6

Y1 - 2012/6

N2 - Dynamic methylations and demethylations of histone lysine residues are important for gene regulation and are facilitated by histone methyltransferases and histone demethylases (HDMs). KDM5B/Jarid1B/PLU1 is an H3K4me3/me2 specific lysine demethylase belonging to the family of JmjC domain containing lysine specific HDMs (JHDMs). Several studies have linked KDM5B to breast, prostate and skin cancer, highlighting its potential as a drug target. However, most inhibitor studies have focused on other JHDMs, and inhibitors for KDM5B remain to be explored. Here, we report the expression, purification and characterization of the catalytic core of recombinant KDM5B (residues 1-769, ccKDM5B). We show that ccKDM5B, recombinantly expressed in insect cells, demethylates H3K4me3 and H3K4me2 in vitro. The kinetic characterization showed that ccKDM5B has a K(m) (app) value of 0.5 µM for its tri-methylated substrate H3(1-15)K4me3, a considerably increased apparent substrate affinity than reported for related histone demethylases. Despite the presence of a PHD domain, the catalytic activity was not affected by additional methylation at the H3K9 position, suggesting that in vitro chromatin cross talk between H3K4 and H3K9 does not occur for ccKDM5B. Inhibition studies of ccKDM5B showed both in vitro and in cell inhibition of ccKDM5B by 2,4-pyridinedicarboxylic acid (2,4-PDCA) with a potency similar to that reported for the histone demethylase KDM4C. Structure-guided sequence alignment indicated that the binding mode of 2,4-PDCA is conserved between KDM4A/C and KDM5B.

AB - Dynamic methylations and demethylations of histone lysine residues are important for gene regulation and are facilitated by histone methyltransferases and histone demethylases (HDMs). KDM5B/Jarid1B/PLU1 is an H3K4me3/me2 specific lysine demethylase belonging to the family of JmjC domain containing lysine specific HDMs (JHDMs). Several studies have linked KDM5B to breast, prostate and skin cancer, highlighting its potential as a drug target. However, most inhibitor studies have focused on other JHDMs, and inhibitors for KDM5B remain to be explored. Here, we report the expression, purification and characterization of the catalytic core of recombinant KDM5B (residues 1-769, ccKDM5B). We show that ccKDM5B, recombinantly expressed in insect cells, demethylates H3K4me3 and H3K4me2 in vitro. The kinetic characterization showed that ccKDM5B has a K(m) (app) value of 0.5 µM for its tri-methylated substrate H3(1-15)K4me3, a considerably increased apparent substrate affinity than reported for related histone demethylases. Despite the presence of a PHD domain, the catalytic activity was not affected by additional methylation at the H3K9 position, suggesting that in vitro chromatin cross talk between H3K4 and H3K9 does not occur for ccKDM5B. Inhibition studies of ccKDM5B showed both in vitro and in cell inhibition of ccKDM5B by 2,4-pyridinedicarboxylic acid (2,4-PDCA) with a potency similar to that reported for the histone demethylase KDM4C. Structure-guided sequence alignment indicated that the binding mode of 2,4-PDCA is conserved between KDM4A/C and KDM5B.

KW - The Faculty of Pharmaceutical Sciences

KW - BRIC

U2 - 10.1111/j.1742-4658.2012.08567.x

DO - 10.1111/j.1742-4658.2012.08567.x

M3 - Journal article

VL - 279

SP - 1905

EP - 1914

JO - F E B S Journal

T2 - F E B S Journal

JF - F E B S Journal

SN - 1742-464X

IS - 11

ER -

ID: 37815750