Please do not adjust margins
ChemComm
Page 4 of 4
DOI: 10.1039/C6CC00095A
COMMUNICATION
Journal Name
Foundation (2013T60613). J.C.E. acknowledges
a
NGS
scholarship. Y.F. is a Wellcome Trust Sir Henry Dale Fellow (WT
100104/Z/12/Z). Y.-T. C. acknowledges funding from the
National Medical Research Council (NMRC/CBRG/0015/2012).
M. V. acknowledges funding from Medical Research Council,
Marie Curie Career Integration Grant (333487) and the WT
Institutional Strategic Support Fund.
References
1
2
I. Fridovich, Annu. Rev. Biochem., 1995, 64, 97.
I. N. Zelko, T. J. Mariani and R. J. Folz, Free Radical Biol. Med.,
2002, 33, 337.
3
4
H. J. Lee, K. J. Korshavn, A. Kochi, J. S. Derrick and M. H. Lim,
Chem. Soc. Rev., 2014, 43, 6672.
M. A. Hough, J. G. Grossmann, S. V. Antonyuk, R. W. Strange,
P. A. Doucette, J. A. Rodriguez, L. J. Whitson, P. J. Hart, L. J.
Hayward, J. S. Valentine and S. S. Hasnain, Proc. Natl. Acad.
Sci. U. S. A., 2004, 101, 5976.
5
6
R. Noor, S. Mittal and J. Iqbal, Med. Sci. Monit., 2002, 8, 210.
R. Gonzales, C. Auclair, E. Voisin, H. Gautero, D. Dhermy and
P. Boivin, Cancer Res., 1984, 44, 4137.
L. Papa, G. Manfredi and D. Germain, Genes and Cancer,
2014, 5, 15.
A. Vasilaki and M. J. Jackson, Free Radical Biol. Med., 2013,
65, 317.
Fig.
5 Fluorogenic response of SODO derivatives upon incubation with serial
7
8
9
concentrations of hCu/Zn SOD in 20 mM Tris-HCl buffer (pH = 7.4). λexc.: 460 nm for
SODO and SODO1, 510 nm for SODO2. ФF in hCu/Zn SOD: SODO: 0.45, SODO1: 0.11,
SODO2: 0.22. Values are represented as means and error bars as standard deviations (n
= 3).
M. Marikovsky, V. Ziv, N. Nevo, C. Harris-Cerruti and O.
Mahler, J. Immunol., 2003, 170, 2993.
10 S. T. Ferreira, L. Stella and E. Gratton, Biophys. J., 1994, 66
1185.
11 E. Luchinat, A. Gianoncelli, T. Mello, A. Galli and L. Banci,
Chem. Commun., 2015, 51, 584.
,
and triazole groups for binding at hCu/Zn SOD (Fig. 5). These
results confirmed the need of four hydrogen bonds, which are
missing in the analogues SODO1 and SODO2 (Fig. S11 in ESI),
to restrict the torsional motion of SODO and induce its 12 D. V. Martyshkin, S. B. Mirov, Y. X. Zhuang, J. P. Crow, V.
Ermilov and J. S. Beckman, Spectrochim. Acta A Mol. Biomol.
Spectrosc., 2003, 59, 3165.
maximal fluorogenic response.
In summary, we have designed a new class of BODIPY
fluorogens with enhanced spectral properties by incorporating
both rotational restriction and PeT-quenching groups. These
new BODIPY fluorogens show excellent properties as polarity
probes with minimal background emission in aqueous media
and long Stokes shifts upon fluorescence activation. In vitro
studies identified one derivative (SODO) as a highly selective
fluorogenic probe for Cu/Zn SOD. SODO shows remarkable
fluorescence emission only after binding to Cu/Zn SOD with
very high selectivity over ROS-related enzymes and
metabolites as well as the other SOD isoforms (i.e. Mn-SOD
and Fe-SOD). The high selectivity of SODO enabled its use for
imaging Cu/Zn SOD in vivo during the onset of an inflammatory
13 P. Shieh, V. T. Dien, B. J. Beahm, J. M. Castellano, T. Wyss-
Coray and C. R. Bertozzi, J. Am. Chem. Soc., 2015, 137, 7145.
14 Y. Hori, T. Norinobu, M. Sato, K. Arita, M. Shirakawa and K.
Kikuchi, J. Am. Chem. Soc., 2013, 135, 12360.
15 J. C. Er, M. K. Tang, C. G. Chia, H. Liew, M. Vendrell and Y.-T.
Chang, Chem. Sci., 2013, 4, 2168.
16 H. Sunahara, Y. Urano, H. Kojima and T. Nagano, J. Am.
Chem. Soc., 2007, 129, 5597.
17 T. Kowada, H. Maeda and K. Kikuchi, Chem. Soc. Rev., 2015,
44, 4953-4972.
18 A. Vázquez-Romero, N. Kielland, M. J. Arévalo, S. Preciado, R.
J. Mellanby, Y. Feng, R. Lavilla and M. Vendrell, J. Am. Chem.
Soc., 2013, 135, 16018.
19 Y.-H. Ahn, J.-S. Lee and Y.-T. Chang, J. Comb. Chem., 2008, 10
376.
,
response in a zebrafish tail fin injury model. Furthermore, we 20 Y. Urano, M. Kamiya, K. Kanda, T. Ueno, K. Hirose and T.
Nagano, J. Am. Chem. Soc., 2005, 127, 4888.
performed computational modelling to analyse the binding of
SODO at Cu/Zn SOD. Structure-activity studies suggest that the
binding occurs at the interface of the two enzymatic subunits
and involves four residues to restrict the torsional motion of
21 W. Qin, V. Leen, T. Rohand, W. Dehaen, P. Dedecker, M. van
der Auweraer, K. Robeyns, L. van Meervelt, D. Beljonne, B.
van Averbeke, J. N. Clifford, K. Driesen, K. Binnemans and N.
l. Boens, J. Phys. Chem. A, 2008, 113, 439.
the BODIPY fluorophore and deactivate its PeT-quenching 22 M. Vendrell, G. G. Krishna, K. K. Ghosh, D. Zhai, J.-S. Lee, Q.
Zhu, Y. H. Yau, S. G. Shochat, H. Kim, J. Chung and Y.-T.
Chang, Chem. Commun., 2011, 47, 8424.
23 J.-S. Lee, M. Vendrell and Y.-T. Chang, Curr. Opin. Chem. Biol.
2011, 15, 760.
24 L. Li, B. Yan, Y. Q. Shi, W. Q. Zhang and Z. L. Wen, J. Biol.
Chem. 2012, 287, 25353.
groups. SODO is the first fluorogenic probe for Cu/Zn SOD and
represents a unique probe for the detection and in vivo
imaging of Cu/Zn SOD during the progression of inflammatory
disorders.
25 J. H. Chen, K. Stoeber, S. Kingsbury, S. E. Ozanne, G. H.
Williams and C. N. Hales, J. Biol. Chem., 2004, 279, 49439.
26 J. A. Tainer, E. D. Getzoff, K. M. Beem, J. S. Richardson and D.
C. Richardson, J. Mol. Biol., 1982, 160, 181.
L.Z. acknowledges the ‘973’ program (2014CB932002), the
Natural Science Foundation of China (11105150) and the
Special Financial Grant from China Postdoctoral Science
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins