Biosensors and Bioelectronics 86 (2016) 68–74
Contents lists available at ScienceDirect
Biosensors and Bioelectronics
journal homepage: www.elsevier.com/locate/bios
A HClO-specific near-infrared fluorescent probe for determination of
Myeloperoxidase activity and imaging mitochondrial HClO in living
cells
Fengshou Tian a,1, Yan Jia b,c,1, Yanan Zhang a,b,c, Wei Song d, Guangjiu Zhao b,c,
Zongjin Qu b,c, Chunyan Li a,b,c, Yahong Chen a, Peng Li b,c,n
a
Department of Chemistry, Zhoukou Normal University, Zhoukou 466001, PR China
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS), 457 Zhongshan
Road, Dalian 116023, PR China
c
Graduate School of the Chinese Academy of Sciences, Beijing 100039, PR China
d
The First Affiliated Hospital of Dalian Medical University, 222 Zhongshan Road, 116011, PR China
b
art ic l e i nf o
a b s t r a c t
Article history:
Received 5 April 2016
Received in revised form
24 May 2016
Accepted 13 June 2016
Available online 14 June 2016
Hyperchlorous acid (HClO), produced from MPO, is recognized as a host defense that kills pathogens; a
signaling molecule that initiates cell apoptosis; and a harmful agent when overproduced. Thus, measuring of endogenous HClO and MPO will always find its great importance in revealing biological roles
under complex biological conditions. In this study, a turn-on near infrared (NIR) fluorescent probe Cy7NphS has been designed and developed for highly selective and sensitive sensing of HClO and Myeloperoxidase (MPO) with fast response time. The newly developed probe has been successfully applied in
real-time monitoring of HClO and MPO activity in PBS solutions and living HL-60 cells. When applied in
MPO activity determination, the probe showed very high sensitivity with a detection limit of as low as
3.69 10 3 U/mL. Furthermore, the living cell imaging study suggested that this probe could detect HClO
in mitochondria.
& 2016 Elsevier B.V. All rights reserved.
Keywords:
Near-infrared
ClO /MPO detection
Bio-imaging
Real-time monitoring
Mitochondria targeting
1. Introduction
Hypochlorous acid (HOCl), a member of the ROS family, was a
strong oxidative agent that played essential roles in many cellular
processes. The most famous physiological function of HClO was to
kill pathogens and thus contribute to host defense (Klebanoff,
2005; Weiss 1989). Recent evidence also suggested that endogenous HClO could serve as a signal to activate caspase and
mediate cell apoptosis (Gloire et al., 2006; Sugiyama et al., 2004).
However, overproduced HClO was believed to cause oxidative injuries to biomolecules such as nucleic acids, proteins and lipids
(Eley et al., 1991; Schraufstatter et al., 1988; Tatsumi and Fliss,
1994). This chemical property of HClO made it a central pathogenic factor in a variety of diseases, such as cardiovascular diseases, atherosclerosis, osteoarthritis, rheumatoid arthritis and lung
n
Corresponding author at: State Key Laboratory of Molecular Reaction Dynamics,
Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS), 457
Zhongshan Road, Dalian 116023, PR China.
E-mail address: [email protected] (P. Li).
1
These authors contributed equally.
http://dx.doi.org/10.1016/j.bios.2016.06.039
0956-5663/& 2016 Elsevier B.V. All rights reserved.
injury (Daugherty et al., 1994; Weitzman and Gordon, 1990;
Winterbourn and Kettle 2000; Wu and Pizzo 2001). Elevated cellular HClO levels could be associated with increased MPO activity,
because HClO was produced by MPO-catalyzed oxidation of Cl- by
H2O2 in cells. It has been pointed out that MPO monitoring allowed identification of patients at risk of cardiac events even in
the absence of myocardial necrosis (Asselbergs et al., 2004; Baldus
et al., 2003; Meuwese et al., 2007). Therefore, the determination of
MPO activity and cellular HClO levels has vast importance in understanding the biological roles of HClO as well as early diagnosis
of HClO-related diseases.
Reaction-based small-molecule fluorescent probes represented
a promising technology for measuring biological species due to its
sensitivity and convenience (Chan et al., 2012; Cho and Sessler,
2009; Li et al., 2014; Yang et al., 2014; Yuan et al., 2013b). Recently,
many efforts have been devoted to the development of fluorescent
probes for measuring HClO (Cheng et al., 2013; Fan et al., 2015;
Hou et al., 2015; Wang et al., 2013; Yue et al., 2015) and MPO
activity (Goiffon et al., 2015; Gross et al., 2009). In this study, our
goal was to develop a HClO-specific fluorescent probe that could
monitor HClO levels and MPO activity in real-time without
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
interference of bioautofluorescence (Ntziachristos et al., 2002;
Yuan et al., 2013a) from the complex biological systems. Development of NIR fluorescent probes for HOCl represents a great
challenge because cyanines, the most commonly used NIR dyes,
are prone to oxidative cleavage and subsequent fluorescence
quenching (Sun et al., 2014; Wang et al., 2009). As an attempt, we
designed and developed probe Cy7-NphS, a new fluorescent probe
that merit HClO selectivity, MPO specificity, high sensitivity, fast
response time, and near-infrared emission at the same time.
Herein we presented the application of Cy7-NphS in determination of MPO activity and imaging of HClO in biological samples.
The results showed that Cy7-NphS was capable of monitoring
MPO-derived HClO in real time in living cells. Through a linear
relationship between the fluorescence intensity and MPO concentrations, the MPO activity could be quantitatively determined
with a detection limit of 3.69 10 3 U/mL. The results also
showed that Cy7-NphS was mitochondria targeted and could image mitochondrial HClO in living cells.
69
1000 μL Acetonitrile and 3985 μL PBS buffers (pH ¼7.40), several
ROS including NaClO was added. The mixture was equilibrated for
30 s before measurement. The fluorescence intensity was measured at λex ¼750 nm. The excitation and emission slits were set
to 5 nm, respectively.
2.4. General procedure for monitoring Myeloperoxidase
Measurement of MPO activity was carried out in 100 mM PBS
(pH 7.40). With a total volume of 3000 μL, culture mixtures that
contained PBS, Cy7-NphS and specified amount of MPO were
mixed gently. Then reactions were started by addition of proper
amount of H2O2. To ensure that metabolites formation was enzyme and H2O2 dependent, control incubations were carried out
without enzyme sources/H2O2 at the same time. The solution was
incubated at 37 °C over 20 min while determined the fluorescent
intensity of λem ¼790 nm (λex ¼ 750 nm ) at 1 min interval.
2.5. Cell culture and confocal fluorescence imaging
2. Experimental section
2.1. Materials and instruments
Common reagents or materials were obtained from commercial
source of analytical reagent grade, and used without further purification except as otherwise noted. MPO Peroxidase (MPO) and
Salicyl hydroxamic acid (SHA) was purchased from Sigma Aldrich.
Ultrapure water was used throughout the analytical experiments.
A series of ROS is obtained or prepared as described in detail in
supporting information. Steady-state UV/Vis was measured at
room temperature on a Lambda 35 UV–visible Spectrophotometer
(Perkin-Elmer) with 1.0-cm quartz cells. Fluorescence emission
spectra were obtained at room temperature on a Fluoromax-4
Spectrofluorometer (Horiba-Jobin Yvon), with a Xenon lamp and
1.0-cm quartz cells.
HL-60 cells lines were obtained from the CAS (Chinese Academy of Sciences) Cell Bank. and cultured in IMDM (Hyclone)
medium supplemented with 20% (v/v) fetal bovine serum (FBS) in
a humidified incubator containing 5% CO2 at 37 °C. The cells in
35 mm 12 mm glass bottom cell culture dishes were set at a
density of 2.5 105/mL. The stock solution of Cy7-NphS in DMSO
(1 mM) was diluted with phosphate buffered saline solution
(100 mM, pH 7.4, 138 mM NaCl) with final concentration of 10 μM.
Confocal fluorescence images (Ex. 635 nm, Em. 700–800 nm) were
observed with Olympus FV1000 confocal laser-scanning microscope with an objective lens ( 100). Hoechst and mitotracker
green were used to co-localize in cell imaging study. (Ex. 405 nm,
Em. 450–550 nm) was set for Hoechst and (Ex. 488 nm, Em. 500–
600 nm) was set for mitotracker green.
3. Results and discussion
2.2. Synthesis and structural characterization of Cy7-NphS
3.1. Design and synthesis of Cy7-NphS
2-[4-Chloro-7-(1-ethyl-3,3-dimethyl(indolin-2-ylidene)] 3,5(propane-1,3-diyl) 1,3,5-heptatrien-1-yl) 1-ethyl-3,3-dimethyl3H-indolium (Cy7-Cl) was synthesized according to a slight
modification of the literature procedure. After that, the synthesis
of Cy7-NphS was described as below. Under the nitrogen atmosphere, a mixture containing Cy7-Cl and 5 equiv of 4-Amino
thioanisole, (C7H9NS) in anhydrous N, N′-dimethylformamide
(DMF) was stirred at 90 °C. The reaction was monitored by TLC
analysis and was stopped when there was no more Cy7-Cl. Then,
the DMF was removed under reduced pressure. The resulting residue was purified on a short-column chromatography (silica gel,
ethyl acetate: methanol¼ 5:1).1HNMR(400 MHz, d6-DMSO) δ
(ppm): 8.70(s, 1 h), 7.93(d, 2 h), 7.44(d, J¼ 7.3 Hz, 2 h), 7.26(m, 6 h),
7.12(t, J ¼7.5 Hz, 2 h), 6.89(d, J¼8.6 Hz, 2 h), 6.04(d, J ¼14.0 Hz,
2 h), 4.08(m, J¼7.0 Hz, 4 h), 2.6(s, 4 h), 2.30(s, 3 h), 1.85(s, 2 h), 1.30
(s, 10 h), 1.22(s, 8 h) 13CNMR (d6-DMSO, 500 MHz) δ(ppm):
170.45, 157.97, 145.98, 142.74, 142.39, 141.28, 130.66, 128.91, 127.53,
124.82, 122.86, 117.33, 110.83, 98.76, 48.70, 38.84, 28.02, 24.70,
21.82, 17.68, 12.42 HRMS: m/z C41H50N3S þ Calcd 614.36, found
[M]þ(614.36).
2.3. General procedure for monitoring ROS
On the basis of the procedure following, all the measurements
of the activity of different ROS were carried out in 100 mM PBS
(pH7.40). The probe Cy7-NphS (Ethanol, 15 μL, 1.0 mM) was added
to a 10.0-mL color comparison tube. After dilution to 3.0 μM with
The development of NIR fluorescent probes for HClO with
turned-on fluorescence represented a great challenge because
cyanines, the most commonly used NIR dyes, were vulnerable to
HClO attack. As a result, the NIR fluorescence of cyanine-based
probes for HClO was usually quenched due to the oxidative cleavage of the cyanine backbone when they responded to this species.
Our Strategy for designing a “turn-on” NIR fluorescent probe for
HClO relied on the facilitate functionalizaion of a heptacyanine dye
with 4-(methylthio)-benzenamine group, a sulfur-containing
moiety that reacted with HClO more kinetically superior than the
cyanine backbone (Scheme 1). The mechanism of the fluorescence
enhancement of CyNPhS in response to HClO can be explained by
photoinduced electron transfer (PET) process (detailed information provided in supporting information). The advantage of this
design could be rationalized as (1) the sulfur-containing moiety as
the HClO reaction center could rule out the concerns of HClO-induced NIR fluorescence quenching; (2) the oxidation of the sulfur
center could be expected as an effective means for modulating the
fluorescence of the cyanine; (3) the designed probe could be
synthesized in a simple one-step reaction. The details of the synthetic procedure of Cy7-NphS are described in the Supporting
information (Scheme S1). The chemical structures of Cy7-NphS
were verified by HRMS, 1H NMR, and 13C NMR (see Figs. S1 3,
ESI†).
After getting this compound, we then tested its fluorescence
response to HClO in the NIR region. As shown in Fig. S4 (ESI†), Cy7-
70
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
Scheme 1. Proposed mechanism of Cy7-NphS responding to HClO and MPO.
NphS showed λmax for absorption and emission at 752 nm and
789 nm, respectively, both of which lie in the NIR region. Upon
addition of sodium hypochlorite (NaOCl) as the HOCl source into
the buffer solution containing Cy7-NphS, the NIR fluorescence
intensity could increase by 20-fold (Fig. 1(a)). The product of the
oxidation reaction of Cy7-NphS by HClO was analyzed by high
resolution mass spectrometry experiments, and new peaks of m/z
þ16 were found (Fig. S5, ESI†), indicating the formation of Cy7NphSO (Scheme 1). These results demonstrated the success of the
design strategy, and promoted us to further explore the feasibility
and practicability of the use of the probe in HClO detection.
3.2. Investigations of spectral properties of Cy7-NphS
Firstly, the selectivity of Cy7-NphS toward HClO over other
reactive oxygen species (ROS) was investigated under
physiological conditions (pH 7.4 at 37 °C). In this case, the fluorescence response of Cy7-NphS to various biological ROS, such as
HClO, H2O2, OH, BuOO , BuOOH, NO and O2- were measured. The
results showed that no significant fluorescence changes were observed at 789 nm except for HClO (Fig. 1(b)). In addition, timedependent experiments were carried out to determine the effect
of reaction time on the selectivity (Fig. S6, ESI†). The results demonstrated the inert reactivity of Cy7-NphS toward ROS other
than HClO regardless of reaction time. The results also showed
that the fluorescence of oxidized Cy7-NphS was sufficiently stable
for fluorescence readout. Importantly, the kinetic curves revealed
that Cy7-NphS responded to HClO extremely fast, with the fluorescence reaching a plateau typically in 30 s. The fast responding
rate constituted the basis for real-time monitoring HClO levels and
MPO activity in biological samples.
Subsequently, the ability of Cy7-NphS in quantitative analysis
of HClO was evaluated. The fluorescence changing of Cy7-NphS
(3 μM) upon addition of NaOCl at different concentrations was
measured after 1 min of mixing (100 mM PBS, pH 7.40). Fig. 1
(c) demonstrated that there was a linear dependence of the
fluorescence intensity on the HClO concentration in the range of
0–3.84 μM, with a regression equation was (R2 ¼0.9982). The detection limit (LOD, 3s/k) and quantification limit (LOQ, 10s/k) for
HClO were estimated to be 0.62 70.09 and 1.86 7 0.09 μM, respectively, which were sensitive enough for determining HClO in
cells (Fig. S7, ESI†).
The pH effects on the fluoresence intensity of Cy7-NphS in the
absence and presence of NaOCl was also investigated. As a result,
the fluorescence intensities at 789 nm of free Cy7-NphS and Cy7NphS in the presence of HClO all remained stable in the pH range
of 6.92–8.48 (Fig. S8, ESI†).
Fig. 1. a. Fluorescence spectra of Cy7-NphS (3.0 μM) in the absence of a series of HClO (0–3.84 μM ) excited at 750 nm. b. Fluorescence responses of Cy7-NphS (10 μM) to
various reactive oxygen species. Bars represent the F789 nm intensity, which responds to various ROS in different concentrations [blank (control)], HClO (15 μM), H2O2
(100 μM), OH (100 μM), BuOO (100 μM), BuOOH (100 μM), NO (200 μM) and O2 (200 μM). The mixture was equilibrated for 30 s before the measurement. c. NaOCl
concentration-dependent fluorescence intensity changes that are determined using Cy7-NphS (3 μM) and NaOCl (0–3.84 μM).
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
71
Fig. 2. a. Fluorescence time dependence of Cy7-NphS (3 μM) upon MPO (0.25 U/mL) and H2O2 (8 μM) addition in PBS 7.4 in the presence of Cl- (2 mM). Negative controls
included only MPO or H2O2. b. Inhibitory effects on the oxidation of Cy7-NphS with use of two selective MPO inhibitors. The spectra were measured in PBS at 37 °C for
20 min. Here, residual activity of MPO is represented by the ratio of Finhibited case/Fnormal case. Residual activity of SHA treated case and 4-AB treated case were 0.05 7 0.01 and
0.0337 0.006.
3.3. In situ monitoring of HClO produced from MPO
As intracellular HClO was produced via MPO-catalyzed peroxidation reactions, it was essential to interrogate whether Cy7NphS could be used for in situ monitoring MPO-derived HClO
(Weiss, 1989). For this purpose, Cy7-NphS was introduced to the
MPO/H2O2/Cl- reaction system and the NIR fluorescence was recorded. Fig. 2(a) showed the time-dependent fluorescence intensity at 789 nm in PBS solutions that contained 2 mM Cl-. When
H2O2 was added to PBS solutions that contained Cy7-NphS and
MPO, an immediate fluorescence increment was observed. The
fluorescence increment lasted for ca. 500 s and then became
steady. This fluorescence time-dependence was consistent with
known MPO-catalyzed HClO production. In contrast, no obvious
fluorescence recovery was observed in the absence of either MPO
or H2O2, ruling out the direct oxidation of Cy7-NphS by these
potential interference. To further confirm that the fluorescence
changes was MPO dependent, chemical inhibition assays were
conducted by using two selective MPO inhibitors. As shown in
Fig. 2(b), the fluorescence increment could be potently inhibited
by salicylhydroxamic acid (Davies and Edwards, 1989; Ikeda-Saito
et al., 1991) (50 μM) and 4-aminobenzoic acid hydrazide (Burner
and Obinger, 1997; Kettle et al., 1995) (10 μM ). These results demonstrated that Cy7-NphS was useful in in situ monitoring MPOderived HClO.
3.4. Quantitative determination of MPO activity
Quantitative determination of MPO activity was clinical significant because it discriminated patients of cardiac disease from
healthy persons (Nicholls and Hazen, 2009; Podrez et al., 2000).
Therefore, we examined the ability of Cy7-NphS in quantitative
determination of MPO activity in vitro. H2O2 (8 μM) was added to
PBS solutions containing 3 μM Cy7-NphS and different concentrations of MPO, and the fluorescence intensity at 789 nm were
measured by a fluorescence spectrometer. As shown Fig. 3, the
emission intensities at 789 nm exhibited an excellent linear relationship (R2 ¼0.9950) to the concentrations of MPO in the range
from 0 to 0.275 U/mL. The detection limit (LOD, 3s/k) and quantification limit (LOQ, 10s/k) for MPO were estimated as
0.002137 0.00075 and 0.00629 7 0.00075 U/mL, respectively (Fig.
S9, ESI†). The detection range and limit perfectly covered the human MPO range, i.e., 0.05 70.06 U/mL for the normal group and
0.14 70.04 U/mL for the sick group (Over et al., 1993).
Fig. 3. MPO concentration-dependent fluorescence intensity changes that are determined using Cy7-NphS (3 μM), H2O2 (8 μM) and MPO.
3.5. Imaging of MPO-derived ClO in HL-60 cells
Fluorescence imaging by confocal fluorescence microscopy allowed for direct visualization of intracellular events (Wilkinson,
1984). Our results demonstrated that Cy7-NphS in combination
with confocal fluorescence microscopy was effective in imaging
intracellular HClO. As shown in Fig. 4, Cy7-NphS loaded HL-60
cells exhibited weak intracellular fluorescence. However, strong
intracellular fluorescence arise in 5 min after the cells were stimulated with H2O2. This fluorescence could become much
stronger after another 10 min. In contrast, neither the control
group nor the 4-AB pretreated cells showed increased intracellular
fluorescence during the overall test time. In addition, the timedependent fluorescence image of intracellular HClO in
H2O2-stimulated cells and the plot of relative fluorescence intensity against time were shown in Figs. S10 and S11, respectively.
The results suggested that Cy7-NphS could serve as an efficient
tool for visualizing intracellular HClO in real time.
3.6. Imaging of Mitochondrial HClO
The subcellular locations of HClO could be a significant factor in
determining its biological roles. In myeloid cells, HClO could be
produced in azurophilic granules as MPO located in these granules
(Lemansky et al., 2003). However, HClO may also play their biological roles in other organelles. Mitochondria were the cellular
power plants. They were also involved in signaling and the control
72
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
Fig. 4. a–i. Fluorescence confocal microscopic images of HL-60 cells loaded with 10 μM Cy7-NphS and exposed to stimulation. Scale bar¼ 20 mm. Images were acquired by
using excitation and emission windows of λex¼635 nm and λem ¼700–800 nm, respectively. (a) Cy7-NphS loaded Cells were pictured at the beginning of H2O2 stimulation.
(b) Cy7-NphS loaded cells treated with nothing at the zero point.(c) Cy7-NphS loaded Cells were pre-incubated with 4-AB for 10 min and then pictured before H2O2
stimulation. (d) Cy7-NphS loaded Cells incubated with H2O2 for 5 min (e) Cy7-NphS loaded Cells incubated for 5 min (f) Cy7-NphS loaded Cells pre-incubated with 4-AB and
then stimulated with H2O2 for 5 min (g) Cy7-NphS loaded Cells incubated with H2O2 for 15 min (h) Cy7-NphS loaded Cells incubated for 15 min (i) Cy7-NphS loaded Cells
pre-incubated with 4-AB and then stimulated with H2O2 for 15 min.
of the cell cycle (Hoye et al., 2008). We next assessed whether
Cy7-NphS could respond to mitochondrial HClO in living cells. Colocalization methods (Nelson et al., 2007) which could describe
the existence of two or more dyes in precisely the same space
were employed to confirm the location of Cy7-NphS in HL-60 cells.
HL-60 cells were co-stained with Cy7-NphS and mitotrackergreen, a mitochondria specific dye (Presley et al., 2003), stimulated
with H2O2, and then subject to fluorescence imaging. As shown in
Fig. 5, image of Cy7-NphS-staining (part a) merged well with the
image of mitotracker-green-staining (part b) in compositing image
(part c). The intensity profiles of the linear regions of interest
across HL-60 cells co-stained with Cy7-NphS and mitotrackergreen are varying in close synchrony, with the Pearson's coefficient Rr ¼0.88 and the Manders’coefficients m1 ¼0.90, m2 ¼0.85
(using Image-Pro Plus software). We also employed this software
to carry a color intensity correlation analysis of parts a and b of
Fig. 5 by plotting the intensity of stain Cy7-NphS against mitotracker-green for each pixel to assess the intensity distribution of
the two co-localization dyes. The dependent staining in parts a
and b resulted in highly correlated plots (parts d and e of Fig. 5).
All of these results indicated that Cy7-NphS was able to detect
HClO in mitochondria.
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
73
Fig. 5. Cy7-NphS and mitotracker-green dye colocalization at mitochondria in HL-60 cells. Cells were treated with H2O2 (10 μM) for 0.5 h. The cells were loaded with 1 μM
Cy7-NphS (a), and 200 nM mitotracker-green (b) for 15 min. Fluorescence confocal microscopic images constructed from 700 to 800 nm for (a) and from 500 to 600 nm for
(b) fluorescence collection windows, λex¼635 and 488 nm, respectively. (c) Merged red channel (a) and green channel (b). (d) Displayed the colocalization areas of the red
and green channels selected. (e) Colocalization plot of (a) and (b), z axis represents the frequency that color pair exists in (c), y and x axis represent the intensity of each pixel
in (d). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
4. Conclusions
Acknowledgment
In summary, we have developed a fluorescence probe Cy7NphS for selective and fast detection of HClO. The probe was designed by direct functionalization of a cyanine by 4-(methylthio)benzenamine group, and exhibited turn-on fluorescence in the NIR
region when responding to HClO. The probe was able to monitor
HClO and the MPO activity in PBS solutions as well as in living HL60 cells, and could be used to quantify MPO activity in a range that
covered healthy and sick people. Co-staining experiments demonstrated that the probe was mitochondrial targeted and could
respond to HClO in mitochondria. These results confirmed that
Cy7-NphS was a viable fluorescence probe for measuring HClO and
MPO activity in biological samples.
This work was supported by the National Natural Science
Foundation of China, China (NSFC Nos. 21422309 and 21573229).
GJZ also thanks the financial support from the Frontier Science
Project of the Knowledge Innovation Program of Chinese Academy
of Sciences (CAS) and the CAS Youth Innovation Promotion
Association.
Appendix A. Supplementary material
Supplementary data associated with this article can be found in
the online version at http://dx.doi.org/10.1016/j.bios.2016.06.039.
References
Author contributions
These authors contributed equally.
Note
The authors declare no competing financial interest.
Asselbergs, F.W., Tervaert, J.W.C., Tio, R.A., 2004. Prognostic value of myeloperoxidase in patients with chest pain. New Engl. J. Med. 350 (5) 517-517.
Baldus, S., Heeschen, C., Meinertz, T., Zeiher, A.M., Eiserich, J.P., Munzel, T., Simoons,
M.L., Hamm, C.W., Investigators, C., 2003. Myeloperoxidase serum levels predict
risk in patients with acute coronary syndromes. Circulation 108 (12),
1440–1445.
Burner, U., Obinger, C., 1997. Transient-state and steady-state kinetics of the oxidation of aliphatic and aromatic thiols by horseradish peroxidase. Febs. Lett.
411 (2–3), 269–274.
Chan, J., Dodani, S.C., Chang, C.J., 2012. Reaction-based small-molecule fluorescent
probes for chemoselective bioimaging. Nat. Chem. 4 (12), 973–984.
74
F. Tian et al. / Biosensors and Bioelectronics 86 (2016) 68–74
Cheng, G.H., Fan, J.L., Sun, W., Sui, K., Jin, X., Wang, J.Y., Peng, X.J., 2013. A highly
specific BODIPY-based probe localized in mitochondria for HClO imaging.
Analyst 138 (20), 6091–6096.
Cho, D.G., Sessler, J.L., 2009. Modern reaction-based indicator systems. Chem. Soc.
Rev. 38 (6), 1647–1662.
Daugherty, A., Dunn, J.L., Rateri, D.L., Heinecke, J.W., 1994. Myeloperoxidase, a
catalyst for lipoprotein oxidation, is expressed in human atherosclerotic lesions.
J. Clin. Invest. 94 (1), 437–444.
Davies, B., Edwards, S.W., 1989. Inhibition of myeloperoxidase by salicylhydroxamic
acid. Biochem. J. 258 (3), 801–806.
Eley, D.W., Eley, J.M., Korecky, B., Fliss, H., 1991. Impairment of cardiac contractility
and sarcoplasmic-reticulum Ca2 þ Atpase activity by hypochlorous acid – reversal by dithiothreitol. Can. J. Physiol. Pharmacol. 69 (11), 1677–1685.
Fan, J.L., Mu, H.Y., Zhu, H., Wang, J.Y., Peng, X.J., 2015. Light up ClO- in live cells using
an aza-coumarin based fluorescent probe with fast response and high sensitivity. Analyst 140 (13), 4594–4598.
Gloire, G., Legrand-Poels, S., Piette, J., 2006. NF-kappa B activation by reactive
oxygen species: fifteen years later. Biochem. Pharmacol. 72 (11), 1493–1505.
Goiffon, R.J., Martinez, S.C., Piwnica-Worms, D., 2015. A rapid bioluminescence
assay for measuring myeloperoxidase activity in human plasma. Nat. Commun.
6, 6271.
Gross, S., Gammon, S.T., Moss, B.L., Rauch, D., Harding, J., Heinecke, J.W., Ratner, L.,
Piwnica-Worms, D., 2009. Bioluminescence imaging of myeloperoxidase activity in vivo. Nat. Med. 15 (4), 455–461.
Hou, J.T., Li, K., Yang, J., Yu, K.K., Liao, Y.X., Ran, Y.Z., Liu, Y.H., Zhou, X.D., Yu, X.Q.,
2015. A ratiometric fluorescent probe for in situ quantification of basal mitochondrial hypochlorite in cancer cells. Chem. Commun. 51 (31), 6781–6784.
Hoye, A.T., Davoren, J.E., Wipf, P., Fink, M.P., Kagan, V.E., 2008. Targeting Mitochondria. Acc. Chem. Res. 41 (1), 87–97.
Ikeda-Saito, M., Shelley, D.A., Lu, L., Booth, K.S., Caughey, W.S., Kimura, S., 1991.
Salicylhydroxamic acid inhibits myeloperoxidase activity. J. Biol. Chem. 266 (6),
3611–3616.
Kettle, A.J., Gedye, C.A., Hampton, M.B., Winterbourn, C.C., 1995. Inhibition of
Myeloperoxidase by benzoic-acid hydrazides. Biochem. J. 308, 559–563.
Klebanoff, S.J., 2005. Myeloperoxidase: friend and foe. J. Leukoc. Biol. 77 (5),
598–625.
Lemansky, P., Gerecitano-Schmidek, M., Das, R.C., Schmidt, B., Hasilik, A., 2003.
Targeting myeloperoxidase to azurophilic granules in HL-60 cells. J. Leukoc.
Biol. 74 (4), 542–550.
Li, X.H., Gao, X.H., Shi, W., Ma, H.M., 2014. Design strategies for water-soluble small
molecular chromogenic and fluorogenic probes. Chem. Rev. 114 (1), 590–659.
Meuwese, M.C., Stroes, E.S.G., Hazen, S.L., van Miert, J.N., Kuivenhoven, J.A., Schaub,
R.G., Wareham, N.J., Luben, R., Kastelein, J.J.P., Khaw, K.T., Boekholdt, S.M., 2007.
Serum Myeloperoxidase levels are associated with the future risk of coronary
artery disease in apparently healthy individuals - The EPIC-Norfolk prospective
population study. J. Am. Coll. Cardiol. 50 (2), 159–165.
Nelson, B.K., Cai, X., Nebenfuhr, A., 2007. A multicolored set of in vivo organelle
markers for co-localization studies in Arabidopsis and other plants. Plant J. 51
(6), 1126–1136.
Nicholls, S.J., Hazen, S.L., 2009. Myeloperoxidase, modified lipoproteins, and
atherogenesis. J. Lipid Res. 50 (Suppl), S346–S351.
Ntziachristos, V., Tung, C.-H., Bremer, C., Weissleder, R., 2002. Fluorescence molecular tomography resolves protease activity in vivo. Nat. Med. 8 (7), 757–761.
Over, C., Yamalik, N., Yavuzyilmaz, E., Ersoy, F., Eratalay, K., 1993. Myeloperoxidase
activity in peripheral blood, neutrophil crevicular fluid and whole saliva of
patients with periodontal disease. J. Nihon. Univ. Sch. Dent. 35 (4), 235–240.
Podrez, E.A., Abu-Soud, H.M., Hazen, S.L., 2000. Myeloperoxidase-generated oxidants and atherosclerosis. Free Radic. Biol. Med 28 (12), 1717–1725.
Presley, A.D., Fuller, K.M., Arriaga, E.A., 2003. MitoTracker Green labeling of mitochondrial proteins and their subsequent analysis by capillary electrophoresis
with laser-induced fluorescence detection. J. Chromatogr. B-Anal. Technol.
Biomed. Life Sci. 793 (1), 141–150.
Schraufstatter, I., Hyslop, P.A., Jackson, J.H., Cochrane, C.G., 1988. Oxidant-induced
DNA damage of target-cells. J. Clin. Invest. 82 (3), 1040–1050.
Sugiyama, S., Kugiyama, K., Aikawa, M., Nakamura, S., Ogawa, H., Libby, P., 2004.
Hypochlorous acid, a macrophage product, induces endothelial apoptosis and
tissue factor expression – involvement of myeloperoxidase-mediated oxidant in
plaque erosion and thrombogenesis. Arter. Thromb. Vasc. Biol. 24 (7),
1309–1314.
Sun, M., Yu, H., Zhu, H., Ma, F., Zhang, S., Huang, D., Wang, S., 2014. Oxidative
cleavage-based near-infrared fluorescent probe for hypochlorous acid detection
and myeloperoxidase activity evaluation. Anal. Chem. 86 (1), 671–677.
Tatsumi, T., Fliss, H., 1994. Hypochlorous acid and chloramines increase endothelial
permeability – possible involvement of cellular zinc. Am. J. Physiol. -Heart Circ.
Physiol. 267 (4), H1597–H1607.
Wang, B.S., Li, P., Yu, F.B., Song, P., Sun, X.F., Yang, S.Q., Lou, Z.R., Han, K.L., 2013. A
reversible fluorescence probe based on Se-BODIPY for the redox cycle between
HClO oxidative stress and H2S repair in living cells. Chem. Commun. 49 (10),
1014–1016.
Wang, M., Holmes-Davis, R., Rafinski, Z., Jedrzejewska, B., Choi, K.Y., Zwick, M.,
Bupp, C., Izmailov, A., Paczkowski, J., Warner, B., Koshinsky, H., 2009. Accelerated photobleaching of a cyanine dye in the presence of a ternary target
DNA, PNA probe, dye catalytic complex: a molecular diagnostic. Anal. Chem. 81
(6), 2043–2052.
Weiss, S.J., 1989. Tissue Destruction by Neutrophils. New Engl. J. Med. 320 (6),
365–376.
Weitzman, S.A., Gordon, L.I., 1990. Inflammation and Cancer - Role of PhagocyteGenerated Oxidants in Carcinogenesis. Blood 76 (4), 655–663.
Wilkinson, J.G., 1984. Principles of fluorescence spectroscopy. In: Lakowicz, Jr,
Chemistry in Britain, vol. 20(5), pp. 442-442.
Winterbourn, C.C., Kettle, A.J., 2000. Biomarkers of myeloperoxidase-derived hypochlorous acid. Free Radic. Biol. Med. 29 (5), 403–409.
Wu, S.M., Pizzo, S.V., 2001. alpha(2)-Macraglobulin from rheumatoid arthritis synovial fluid: functional analysis defines a role for oxidation in inflammation.
Arch. Biochem. Biophys. 391 (1), 119–126.
Yang, Z.G., Cao, J.F., He, Y.X., Yang, J.H., Kim, T., Peng, X.J., Kim, J.S., 2014. Macro-/
micro-environment-sensitive chemosensing and biological imaging. Chem. Soc.
Rev. 43 (13), 4563–4601.
Yuan, L., Lin, W.Y., Zheng, K.B., Zhu, S.S., 2013b. FRET-based small-molecule fluorescent probes: rational design and bioimaging applications. Acc. Chem. Res. 46
(7), 1462–1473.
Yuan, L., Lin, W., Zheng, K., He, L., Huang, W., 2013a. Far-red to near infrared analyte-responsive fluorescent probes based on organic fluorophore platforms for
fluorescence imaging. Chem. Soc. Rev. 42 (2), 622–661.
Yue, Y.K., Huo, F.J., Yin, C.X., Chao, J.B., Zhang, Y.B., Wei, X., 2015. An ICT based ultraselective and sensitive fluorescent probe for detection of HClO in living cells.
RSC Adv. 5 (95), 77670–77672.
本文献由“学霸图书馆-文献云下载”收集自网络，仅供学习交流使用。
学霸图书馆（www.xuebalib.com）是一个“整合众多图书馆数据库资源，
提供一站式文献检索和下载服务”的24 小时在线不限IP 图书馆。
图书馆致力于便利、促进学习与科研，提供最强文献下载服务。
图书馆导航：
图书馆首页
文献云下载
图书馆入口
外文数据库大全
疑难文献辅助工具