A photoreactive small-molecule probe for 2-oxoglutarate oxygenases

Dante Rotili, Mikael Altun, Akane Kawamura, Alexander Wolf, Roman Fischer, Ivanhoe K H Leung, Muhammad Mukram Mohamed Mackeen, Ya Min Tian, Peter J. Ratcliffe, Antonello Mai, Benedikt M. Kessler, Christopher J. Schofield

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in human crude cell extracts, including to proteins at endogenous levels. This approach is useful for inhibitor profiling, as demonstrated by crosslinking to the histone demethylase FBXL11 (KDM2A) in HEK293T nuclear extracts. The results also suggest that small-molecule probes may be suitable for substrate identification studies.

Original languageEnglish
Pages (from-to)642-654
Number of pages13
JournalChemistry and Biology
Volume18
Issue number5
DOIs
Publication statusPublished - 27 May 2011
Externally publishedYes

Fingerprint

Oxygenases
Molecules
Crosslinking
Histone Demethylases
Hydroxyquinolines
Cell Extracts
Complex Mixtures
Recombinant Proteins
Purification
Anemia
alpha-ketoglutaric acid
Substrates
Neoplasms
Proteins
Therapeutics

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Molecular Biology
  • Clinical Biochemistry
  • Molecular Medicine
  • Pharmacology

Cite this

Rotili, D., Altun, M., Kawamura, A., Wolf, A., Fischer, R., Leung, I. K. H., ... Schofield, C. J. (2011). A photoreactive small-molecule probe for 2-oxoglutarate oxygenases. Chemistry and Biology, 18(5), 642-654. https://doi.org/10.1016/j.chembiol.2011.03.007

A photoreactive small-molecule probe for 2-oxoglutarate oxygenases. / Rotili, Dante; Altun, Mikael; Kawamura, Akane; Wolf, Alexander; Fischer, Roman; Leung, Ivanhoe K H; Mohamed Mackeen, Muhammad Mukram; Tian, Ya Min; Ratcliffe, Peter J.; Mai, Antonello; Kessler, Benedikt M.; Schofield, Christopher J.

In: Chemistry and Biology, Vol. 18, No. 5, 27.05.2011, p. 642-654.

Research output: Contribution to journalArticle

Rotili, D, Altun, M, Kawamura, A, Wolf, A, Fischer, R, Leung, IKH, Mohamed Mackeen, MM, Tian, YM, Ratcliffe, PJ, Mai, A, Kessler, BM & Schofield, CJ 2011, 'A photoreactive small-molecule probe for 2-oxoglutarate oxygenases', Chemistry and Biology, vol. 18, no. 5, pp. 642-654. https://doi.org/10.1016/j.chembiol.2011.03.007
Rotili D, Altun M, Kawamura A, Wolf A, Fischer R, Leung IKH et al. A photoreactive small-molecule probe for 2-oxoglutarate oxygenases. Chemistry and Biology. 2011 May 27;18(5):642-654. https://doi.org/10.1016/j.chembiol.2011.03.007
Rotili, Dante ; Altun, Mikael ; Kawamura, Akane ; Wolf, Alexander ; Fischer, Roman ; Leung, Ivanhoe K H ; Mohamed Mackeen, Muhammad Mukram ; Tian, Ya Min ; Ratcliffe, Peter J. ; Mai, Antonello ; Kessler, Benedikt M. ; Schofield, Christopher J. / A photoreactive small-molecule probe for 2-oxoglutarate oxygenases. In: Chemistry and Biology. 2011 ; Vol. 18, No. 5. pp. 642-654.
@article{581c79d13c6e4b1998b3c08caf76bd76,
title = "A photoreactive small-molecule probe for 2-oxoglutarate oxygenases",
abstract = "2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in human crude cell extracts, including to proteins at endogenous levels. This approach is useful for inhibitor profiling, as demonstrated by crosslinking to the histone demethylase FBXL11 (KDM2A) in HEK293T nuclear extracts. The results also suggest that small-molecule probes may be suitable for substrate identification studies.",
author = "Dante Rotili and Mikael Altun and Akane Kawamura and Alexander Wolf and Roman Fischer and Leung, {Ivanhoe K H} and {Mohamed Mackeen}, {Muhammad Mukram} and Tian, {Ya Min} and Ratcliffe, {Peter J.} and Antonello Mai and Kessler, {Benedikt M.} and Schofield, {Christopher J.}",
year = "2011",
month = "5",
day = "27",
doi = "10.1016/j.chembiol.2011.03.007",
language = "English",
volume = "18",
pages = "642--654",
journal = "Cell Chemical Biology",
issn = "2451-9448",
publisher = "Elsevier Inc.",
number = "5",

}

TY - JOUR

T1 - A photoreactive small-molecule probe for 2-oxoglutarate oxygenases

AU - Rotili, Dante

AU - Altun, Mikael

AU - Kawamura, Akane

AU - Wolf, Alexander

AU - Fischer, Roman

AU - Leung, Ivanhoe K H

AU - Mohamed Mackeen, Muhammad Mukram

AU - Tian, Ya Min

AU - Ratcliffe, Peter J.

AU - Mai, Antonello

AU - Kessler, Benedikt M.

AU - Schofield, Christopher J.

PY - 2011/5/27

Y1 - 2011/5/27

N2 - 2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in human crude cell extracts, including to proteins at endogenous levels. This approach is useful for inhibitor profiling, as demonstrated by crosslinking to the histone demethylase FBXL11 (KDM2A) in HEK293T nuclear extracts. The results also suggest that small-molecule probes may be suitable for substrate identification studies.

AB - 2-oxoglutarate (2-OG)-dependent oxygenases have diverse roles in human biology. The inhibition of several 2-OG oxygenases is being targeted for therapeutic intervention, including for cancer, anemia, and ischemic diseases. We report a small-molecule probe for 2-OG oxygenases that employs a hydroxyquinoline template coupled to a photoactivable crosslinking group and an affinity-purification tag. Following studies with recombinant proteins, the probe was shown to crosslink to 2-OG oxygenases in human crude cell extracts, including to proteins at endogenous levels. This approach is useful for inhibitor profiling, as demonstrated by crosslinking to the histone demethylase FBXL11 (KDM2A) in HEK293T nuclear extracts. The results also suggest that small-molecule probes may be suitable for substrate identification studies.

UR - http://www.scopus.com/inward/record.url?scp=79957487715&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957487715&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2011.03.007

DO - 10.1016/j.chembiol.2011.03.007

M3 - Article

C2 - 21609845

AN - SCOPUS:79957487715

VL - 18

SP - 642

EP - 654

JO - Cell Chemical Biology

JF - Cell Chemical Biology

SN - 2451-9448

IS - 5

ER -