Please d Co h ne omt Ca do j mu s mt margins
Page 4 of 4
COMMUNICATION
ChemComm
In conclusion, 7OHCou-AACF3, a fluorogenic inhibitor of cPLA with
DOI: 10.1039/C6CC09305A
dual functions of imaging and inhibition has been developed.
7
OHCou-AACF3 has an IC50 of 12.5±1.0 μM which is comparable
with AACOCF . It is able to quench iNOS production via its role as an
3
inhibitor and is capable of imaging the increase in cPLA protein
2
levels. We have also developed Flu7OHCou, a cPLA -selective
2
fluorogenic substrate and demonstrated its use for inhibitor
screening assays.
The authors thank the Ministry of Education (Singapore) (MoE Tier
2
: R-143-000-589-112; YL) and NMRC (1222-2009; CML) for the
financial support and the National University of Singapore for a
Ph.D. scholarship to C.Y.N.
Notes and references
1
2
3
4
5
6
K. Yui, G. Imataka, H. Nakamura, N. Ohara and Y. Naito, Curr.
Neuropharmacol., 2015, 13(6), 776.
M. A. Gijon, D. M. Spencer, A. L. Kaiser and C. C. Leslie, J. Cell
Biol.,1999, 145, 1219.
E. A. Dennis, J. Cao, Y. H. Hsu, V. Magrioti and G. Kokotos,
Chem. Rev., 2011, 111, 6130.
S. A. Farber, E. S. Olson, I. D. Clark and M. E. Halpern, J. Biol.
Chem., 1999, 274, 19338.
J. V. Bonventre, Z. Huang, M. R. Taheri, E. O’Leary, E. Li, M. A.
Moskowitz and A. Sapirstein, Nature, 1997, 390, 622.
C. Desbène, C. Malaplate-Armand, I. Youssef, P. Garcia, C.
Stenger, M. Sauvée, N Fischer, D. Rimet, V. Koziel, M. C.
Escanyé, T. Oster, B. Kriem, F. T. Yen, T. PIllot and J. L. Olivier,
Neurobiol. Aging, 2012, 33, 1123.e17.
7
8
9
M. G. Chiorazzo, N. B. Bloch, A. V. Popov and E. J. Delikatny,
Bioconjugate Chem., 2015, 26, 2360.
W. Huang, A. Bhavsar, R. E. Ward, J. C. E. Hall, J. V. Priestley
and A. T. Michael-Titus, J. Neurotrauma, 2009, 26(8), 1429.
D. Y. Chuang, A. Simonyi, P. T. Kotzbauer, Z. Gu and G. Y. Sun,
J. Neuroinflammation, 2015, 12, 199.
1
1
0 C. S-H Tan, Y-K Ng and W-Y Ong, Mol. Neurobiol., 2016, 53
854.
1 D. Stephens, E. Barbayianni, V. Constantinou-Kokotou, A.
,
3
Peristeraki, D. A. Six, J. Cooper, R. Harkewicz, R. A. Deems, E.
A. Dennis and G. Kokotos, J. Med. Chem., 2006, 49, 2821.
2 D. A. Six, E. Barbayianni, V. Loukas, V. Constantinou-Kokotou,
D. Hadjipavlou-Litina, D. Stephens, A. C. Wong, V. Magrioti,
P. Moutevelis-Minakakis, S. F. Baker, E. A. Dennis and G.
Kokotos, J. Med. Chem., 2007, 50, 4222.
1
1
3 D. Manna and W. Cho, Methods Enzymol., 2007, 434, 15.
Figure 4. Flu7OHCou substrate assay (A) Addition of bee venom
14 S. Connolly, C. Bennion, S. Botterell, P. J. Croshaw, C. Hallam,
K. Hardy, P. Hartopp, C. G. Jackson, S. J. King, L. Lawrence, A.
Mete, D. Murray, D. H. Robinson, G. M. Smith, L. Stein, I.
Walters, E. Wells and W. J. Withnall, J. Med. Chem., 2002, 45
(
5 units/mL) to 20 μM Flu7OHCou constituted in a liposome formed
by vortexing DOPG and DOPC in a Tris-HCl buffer, pH 7.3 with 1%
Triton X-100. Donor peak increased significantly as time increased
from 0min to 60min while acceptor peak decreased with time. (B)
,
1
348
1
5 T. Fukuda, D. K. Kim, M-R. Chin, C. A. Hales and J. V.
Addition of recombinant cPLA (0.05 unit/mL) to 20 μM Flu7OHCou
Bonventre, Am. J. Physiol. Lung Cell Mol. Physiol., 1999, 277,
2
5
33
constituted in a liposome formed by vortexing DOPG and DOPC in a
Tris-HCl buffer, pH 8.9 (i) with 1% Triton X-100, (ii) without Triton X-
1
6 E. A. Dennis, L. J. Reynolds and Y. Lin, Assay and substrate for
arachidonoyl-specific phospholipase A2, Patent
WO1995005479A1, 1995
1
2
00 (C) Addition of recombinant cPLA and sPLA (0.05 unit/mL) to
2 2
0 μM Flu7OHCou constituted in a liposome formed by vortexing
DOPG and DOPC in a Tris-HCl buffer, pH 8.9 (i) without Triton X-100
ii) with 1% Triton X-100.
(
4
Chem. Commun., 2017, 00, 1-3
This journal is © The Royal Society of Chemistry 2017
Please do not adjust margins