PMID- 23219719
OWN - NLM
STAT- MEDLINE
DCOM- 20130828
LR  - 20211021
IS  - 1096-0309 (Electronic)
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 434
IP  - 2
DP  - 2013 Mar 15
TI  - Forster resonance energy transfer competitive displacement assay for human 
      soluble epoxide hydrolase.
PG  - 259-68
LID - S0003-2697(12)00595-7 [pii]
LID - 10.1016/j.ab.2012.11.015 [doi]
AB  - The soluble epoxide hydrolase (sEH), responsible for the hydrolysis of various 
      fatty acid epoxides to their corresponding 1,2-diols, is becoming an attractive 
      pharmaceutical target. These fatty acid epoxides, particularly 
      epoxyeicosatrienoic acids (EETs), play an important role in human homeostatic and 
      inflammation processes. Therefore, inhibition of human sEH, which stabilizes EETs 
      in vivo, brings several beneficial effects to human health. Although there are 
      several catalytic assays available to determine the potency of sEH inhibitors, 
      measuring the in vitro inhibition constant (K(i)) for these inhibitors using 
      catalytic assay is laborious. In addition, k(off), which has been recently 
      suggested to correlate better with the in vivo potency of inhibitors, has never 
      been measured for sEH inhibitors. To better measure the potency of sEH 
      inhibitors, a reporting ligand, 
      1-(adamantan-1-yl)-3-(1-(2-(7-hydroxy-2-oxo-2H-chromen-4-yl)acetyl) 
      piperidin-4-yl)urea (ACPU), was designed and synthesized. With ACPU, we have 
      developed a Forster resonance energy transfer (FRET)-based competitive 
      displacement assay using intrinsic tryptophan fluorescence from sEH. In addition, 
      the resulting assay allows us to measure the K(i) values of very potent compounds 
      to the picomolar level and to obtain relative k(off) values of the inhibitors. 
      This assay provides additional data to evaluate the potency of sEH inhibitors.
CI  - Copyright (c) 2012 Elsevier Inc. All rights reserved.
FAU - Lee, Kin Sing Stephen
AU  - Lee KS
AD  - Department of Entomology and UCD Comprehensive Cancer Center, University of 
      California, Davis, CA 95616, USA.
FAU - Morisseau, Christophe
AU  - Morisseau C
FAU - Yang, Jun
AU  - Yang J
FAU - Wang, Peng
AU  - Wang P
FAU - Hwang, Sung Hee
AU  - Hwang SH
FAU - Hammock, Bruce D
AU  - Hammock BD
LA  - eng
GR  - R01 ES002710/ES/NIEHS NIH HHS/United States
GR  - ES002710/ES/NIEHS NIH HHS/United States
GR  - P42 ES004699/ES/NIEHS NIH HHS/United States
GR  - P42 ES04699/ES/NIEHS NIH HHS/United States
GR  - U54 NS079202/NS/NINDS NIH HHS/United States
GR  - U54 NS079202-01/NS/NINDS NIH HHS/United States
GR  - R37 ES002710/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20121205
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Enzyme Inhibitors)
RN  - 8DUH1N11BX (Tryptophan)
RN  - EC 3.3.2.- (Epoxide Hydrolases)
SB  - IM
MH  - Binding, Competitive
MH  - Biological Assay/*methods
MH  - Crystallography, X-Ray
MH  - Enzyme Activation/drug effects
MH  - Enzyme Inhibitors/chemistry/pharmacology
MH  - Epoxide Hydrolases/antagonists & inhibitors/chemistry/*metabolism
MH  - Fluorescence
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Humans
MH  - Inhibitory Concentration 50
MH  - Solubility
MH  - Tryptophan/chemistry
PMC - PMC3632402
MID - NIHMS427337
EDAT- 2012/12/12 06:00
MHDA- 2013/08/29 06:00
PMCR- 2014/03/15
CRDT- 2012/12/11 06:00
PHST- 2012/09/08 00:00 [received]
PHST- 2012/11/15 00:00 [revised]
PHST- 2012/11/25 00:00 [accepted]
PHST- 2012/12/11 06:00 [entrez]
PHST- 2012/12/12 06:00 [pubmed]
PHST- 2013/08/29 06:00 [medline]
PHST- 2014/03/15 00:00 [pmc-release]
AID - S0003-2697(12)00595-7 [pii]
AID - 10.1016/j.ab.2012.11.015 [doi]
PST - ppublish
SO  - Anal Biochem. 2013 Mar 15;434(2):259-68. doi: 10.1016/j.ab.2012.11.015. Epub 2012 
      Dec 5.