From the Pulmonary and Critical Care Division, Department of Medicine, New England Medical Center/Tupper Research Institute, Tufts University School of Medicine, Boston, Massachusetts 02111
Received for publication, January 17, 2001, and in revised form, January 31, 2001
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ABSTRACT |
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The mechanisms by which ligand-stimulated
generation of reactive oxygen species in nonphagocytic cells mediate
biologic effects are largely unknown. The profibrotic cytokine,
transforming growth factor- Tissue repair responses to injurious agents are complex
processes involving resident/structural (e.g. fibroblasts)
and circulating/immune (e.g. macrophages) cells. A
"normal" repair response most often restores tissue
architecture and maintains organ function. However, in some cases
"dysregulated" repair leads to tissue fibrosis and organ failure.
Cell-cell communication in this context is mediated by a network of
growth factors, cytokines, and other soluble mediators (1-3).
Transforming growth factor- Multiple actions of TGF- We have reported previously on the ability of TGF- The precise function of TGF- Based on the availability of heme peroxidases (such as
phagocyte-derived myeloperoxidases) in the extracellular milieu of "activated" fibroblasts, we postulated that extracellular
H2O2 production by these cells may mediate
similar reactions involving the phenolic amino acid
L-tyrosine. In this study, we examined the ability of
TGF- Cell Culture and Reagents--
All cell culture experiments were
performed on normal human fetal lung fibroblasts (IMR-90; Institute for
Medical Research, Camden, NJ). Cells were plated on 35-mm Petri dishes
at a density of 105 cells/dish and incubated in 5%
CO2, 95% air. For fluoroscopy-based assays, cells were
plated on microscope glass slides sterilely placed in Petri dishes.
Cells were maintained in medium consisting of RPMI 1640 medium (Life
Technologies, Inc.) supplemented with 10% fetal calf serum (Sigma),
100 units/ml penicillin/streptomycin (Sigma), and fungizone (Life
Technologies, Inc.). Medium was changed every 3 days. Cells were grown
to 80-100% confluency and serum-deprived for 48 h prior to
assays. Porcine platelet-derived TGF- Measurement of Dityrosine Formation--
Dityrosine formation
was measured by a fluorometric assay described by Heinecke et
al. (30) with minor modifications. The feasibility and reliability
of this method in our cell culture system was determined first. Known
concentrations of H2O2 (0-3 µM)
were added to a reaction mixture containing 1 mM
L-tyrosine, 5 units/ml horseradish peroxidase (HRP), and 1 mM HEPES, pH 7.4, in Hanks' balanced salt solution (HBSS).
After a 5-min incubation, the reaction was terminated by the addition
of 0.1 M glycine-NaOH buffer, pH 12.0, and the final pH was
adjusted to 10.0. Fluorescence of the samples was monitored at
excitation and emission wavelengths of 325 and 410, respectively.
Values were recorded as relative fluorescence with the sample buffer
containing no H2O2 as reference ("0" value,
Fig. 2, inset).
For dityrosine measurements in cell culture, cells were washed first
with Dulbecco's phosphate-buffered saline and incubated with the same
HBSS-based reaction mixture containing L-tyrosine and HRP
for 1 h at 37 °C. The overlying medium was then removed, the pH
was adjusted to 10.0 with 0.1 M glycine-NaOH buffer, and fluorescence was measured as described above. Samples with the reaction
mixture alone (without cells) that were processed similarly served as
control and the reference against which the relative fluorescence for
each of the experimental samples was recorded. "Spontaneous"
dityrosine formation was not observed in these control samples.
Labeling of Endogenous Proteins Targeted by
H2O2/HRP-mediated Oxidative Tyrosine
Cross-linking--
To detect tyrosine cross-linking on endogenous
protein substrates, we utilized a commercially available laboratory
reagent designed for signal amplification protocols. This reagent,
tyramide, labeled with fluorescein (tyramide-FITC, TSATM Fluorescence
Systems, PerkinElmer Life Sciences), is distributed as "TSA" kits
for amplification of HRP-generated signals in immunoblotting,
immunohistochemistry, and in situ hybridization assays (31).
Specifically, HRP, in the presence of exogenously added
H2O2 (found in the "amplification diluent"
provided with the TSA kit), catalyzes the "cross-linking" of
tyramide-FITC with tyrosine residues of proteins in close proximity to
the site of enzyme (HRP) catalysis. This reportedly results in the
deposition of the fluorophore at or near the site of the bound
antibody-HRP conjugate resulting in "amplification" of the signal
(31). An important feature of this assay is the absolute requirement
for exogenous H2O2 to mediate these
HRP-catalyzed cross-linking reactions. By eliminating exogenous
H2O2 from the reaction, we have exploited this
property of tyramide-FITC to localize "susceptible" tyrosine
residues on cellular or extracellular proteins to mediate similar
cross-linking reactions in response to endogenous
cytokine-generated H2O2 in cultured lung fibroblasts.
Cultured cells grown on Petri dishes were washed first with HBSS and
incubated with 1 ml of HBSS containing 1 mM HEPES buffer (pH 7.4), 5 units/ml HRP, and 1:1000 dilution of freshly reconstituted tyramide-FITC. Tyramide-FITC was supplied by the manufacturer as
lyophilized powder (weight not indicated). Initial reconstitution was
with Me2SO using the 1:10 volume recommended by the
manufacturer. The final concentration of tyramide-FITC used in our
assays amounted to one-half that recommended for use in signal
amplification protocols (kit insert, PerkinElmer Life Sciences). The
amplification diluent (which contains H2O2)
provided with the TSA kit was discarded and is not required for this
application. Cells were allowed to incubate for 1 h at 37 °C in
the buffer solution containing tyramide-FITC and HRP. Cells then were
washed two times with HBSS and visualized (and photographed) with a
fluorescent microscope (Zeiss IM 35 inverted microscope, Oberkochen, Germany).
Statistical Analysis--
Data from the various groups were
expressed as means ± S.D. Statistical comparisons were made using
the Student's t test for unpaired samples. For studies
involving more than two groups, two-way analysis of variance was
determined using the Scheffe's test (GB-STAT, Dynamic Microsystems,
Silver Spring, MD). Statistical significance in all cases was defined
as p < 0.05.
TGF-
To determine then whether physiological concentrations of
H2O2 generated by cytokine-stimulated
fibroblasts are capable of inducing L-tyrosine
dimerization, the same assay medium (with and without HRP) was added to
TGF- Detection and Localization of Endogenous Protein Cross-linking
Reactions Using a Fluorophore-labeled Phenolic Compound
(Tyramide)--
Given the finding that the TGF- Effect of Exogenous H2O2 on the Pattern of
Protein Cross-linking in Non-H2O2-producing
Cells--
The effect of adding H2O2
exogenously to control cells (not treated with TGF- Growth factors and cytokines are now well recognized to induce the
generation of ROS in a variety of nonphagocytic cells for the purposes
of cell signaling and in regulation of normal physiological processes
(reviewed in Ref. 33). The biological effects of ROS are likely to
depend on several factors including the specific reactive species
(e.g. O In this study, we show that "physiological" concentrations of
H2O2 generated by TGF- Similar to arteriosclerosis, pulmonary fibrosis also has been
characterized as a disease of oxidative stress (37), although the
mechanisms by which oxidants might contribute to disease pathogenesis are not known (38). The data presented here suggest one possible, and
even likely, mechanism. The strong association of TGF- The specific ECM protein(s) targeted by
H2O2/HRP-dependent cross-linking
reactions induced by TGF- Whether oxidative cross-linking reactions of the type discussed may
occur under physiologic rather than pathologic conditions is unclear.
Physiologic cross-linking of collagen and elastin is mediated by
lysyl oxidase, a copper amine oxidase that modifies the 1 (TGF-
1), generates
extracellular hydrogen peroxide (H2O2) in contrast to intracellular
reactive oxygen species production by certain mitogenic growth factors
in human lung fibroblasts. To determine whether tyrosine residues in
fibroblast-derived extracellular matrix (ECM) proteins may be targets
of H2O2-mediated
dityrosine-dependent cross-linking reactions in response to
TGF-
1, we utilized fluorophore-labeled tyramide, a structurally
related phenolic compound that forms dimers with tyrosine, as a probe
to detect such reactions under dynamic cell culture conditions. With
this approach, a distinct pattern of fluorescent labeling that seems to
target ECM proteins preferentially was observed in TGF-
1-treated
cells but not in control cells. This reaction required the presence of
a heme peroxidase and was inhibited by catalase or diphenyliodonium (a
flavoenzyme inhibitor), similar to the effect on TGF-
1-induced
dityrosine formation. Exogenous addition of
H2O2 to control cells that do not release
extracellular H2O2 produced a similar
fluorescent labeling reaction. These results support the concept that,
in the presence of heme peroxidases in vivo,
TGF-
1-induced H2O2 production by fibroblasts
may mediate oxidative dityrosine-dependent cross-linking of
ECM protein(s). This effect may be important in the pathogenesis
of human fibrotic diseases characterized by overexpression/activation
of TGF-
1.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
1
(TGF-
1)1 is a
multifunctional cytokine that plays a central role in tissue injury/repair. Persistent or exaggerated expression and/or activation of TGF-
1 in areas of active injury/repair is linked to a number of
human fibrotic diseases involving diverse organ systems (4, 5).
1 contribute to its profibrotic effects
including its ability to transform fibroblasts both in vitro and in vivo into activated contractile phenotypes known as
"myofibroblasts" (6-8). Myofibroblasts are key effector cells in
injury/repair processes and fibrosis because of their high synthetic
capacity for extracellular matrix (ECM) proteins (9, 10), integrins (11), growth factors (12), growth factor receptors (13), and oxidants
(14). Recent studies suggest that TGF-
1-induced myofibroblast
differentiation depends on the compliance/deformability of the matrix
on which cells are grown (15, 16). This observation is consistent with
the growing recognition that biomechanical properties of the ECM
regulate cell function (17-19).
1 to induce
extracellular release of H2O2 in
association with cell surface-associated NADH:flavin:O2
oxidoreductase (NADH oxidase) activity in human lung fibroblasts (14).
The enzymatic source and regulation of this activity is unrelated to
the p21ras-dependent generation of
intracellular reactive oxygen species (ROS) by certain
mitogenic growth factors (20). Intracellular ROS have been reported to
function as signaling molecules in growth factor-induced mitogenesis
(21-23). Recent work by Barrett et al. (24) suggests that
this effect may be mediated by intracellular O
1-induced extracellular
H2O2 production is yet to be determined. Our
previous data indicate that although tyrosine phosphorylation regulates
NADH oxidase activity/H2O2 production,
H2O2 does not itself seem to mediate protein
tyrosine phosphorylation in cultured lung fibroblasts (25). Moreover, TGF-
1-induced extracellular H2O2 is not
associated with cell proliferation in these cells (20).
H2O2 is a relatively mild oxidant in most
biological systems, but it functions as a potent oxidant (by inducing
dimerization) of phenolic compounds in the presence of heme peroxidases
(26). This property of H2O2 is utilized for
useful purposes in the formation of a protective coat, composed of
highly cross-linked ECM proteins, around freshly fertilized oocytes in
sea urchins and in the plant hypersensitivity response (27, 28).
Heinecke et al. (29) have demonstrated that
phagocyte-derived myeloperoxidases, which normally convert H2O2 to hypochlorous acid, are capable of
catalyzing similar reactions in human atherosclerotic plaques.
1 to induce dimerization of exogenous L-tyrosine by
H2O2/heme peroxidase-dependent
oxidation. Moreover, the possibility that this reaction may target
endogenous protein targets under dynamic conditions in which cultured
fibroblasts are actively generating extracellular
H2O2 was assessed utilizing a novel fluorescent labeling approach that identifies susceptible tyrosine residues.
EXPERIMENTAL PROCEDURES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
1 was obtained from R&D Systems
(Minneapolis, MN). All other reagents including L-tyrosine
were from Sigma.
RESULTS
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
1 Induces Dimerization of Exogenous L-Tyrosine
by an H2O2- and Heme
Peroxidase-dependent Mechanism--
The ability of
H2O2 in the presence of heme peroxidases such
as HRP to induce oxidative dimerization of phenolic compounds is well
established (Fig. 1a; see Ref.
26 for details). We hypothesized that TGF-
1, by stimulating
H2O2 production in lung fibroblasts, may
mediate a similar effect on the phenolic amino acid,
L-tyrosine. First, the feasibility of measuring dityrosine
formation in our cell culture system using the method described by
Heinecke et al. (30) was verified by adding increasing
concentrations of H2O2 (0-3 µM)
to assay medium containing L-tyrosine (1 mM)
and HRP (5 units/ml) for 5 min followed by an assessment of
dityrosine formation (see "Experimental Procedures" for details). A
dose-dependent linear relationship between
H2O2 concentration and dityrosine formation was
observed (Fig. 2, inset). When
HRP or L-tyrosine was excluded from the
reaction, dityrosine formation was not detectable (results not
shown).
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Fig. 1.
a, dimerization of substituted phenolic
compounds by heme peroxidase-catalyzed oxidation by
H2O2. The ability of
H2O2 to induce heme peroxidase-catalyzed
dimerization of substituted phenolic compounds is well recognized. This
effect is mediated by a series of reactions that begins with the
reversible oxidization of HRP by H2O2 to an
intermediate compound capable of reacting with phenolic substrates to
form radicals that combine to form dimers (see Ref. 26 for
details). R', alkyl groups. b, structural
comparison of the p-substituted phenolic compounds, tyrosine
and tyramide. Both compounds are capable of undergoing homodimerization
and heterodimerization reactions by the mechanism described in
a. R', fluorescein moiety in tyramide-FITC.
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Fig. 2.
Dityrosine formation by
TGF- 1-induced H2O2
production in the presence of a heme peroxidase, HRP, in cultured human
lung fibroblasts. Inset, increasing
H2O2 concentrations were added to assay medium
containing HRP (5 units/ml) and L-tyrosine (1 mM) for 5 min, and dityrosine formation was measured as
described under "Experimental Procedures." Using this assay system,
the effect of endogenous H2O2 production on
dityrosine formation was assessed in control and TGF-
1-stimulated (2 ng/ml × 16 h) fibroblasts in the presence/absence of
HRP (5 units/ml), catalase (1000 units/ml), or
diphenyliodonium (DPI, 10 µM, a flavoprotein
inhibitor added 15 min prior to assay). The starting
concentration of L-tyrosine in all cases was 1 mM. Values of dityrosine formation are expressed as
mean ± SD, n = 4.
1-treated fibroblasts at the peak time point when these cells
have been shown to actively release H2O2 into
the overlying media (14). In the presence of HRP, dityrosine formation
was stimulated markedly in TGF-
1-treated cells, whereas it was
undetectable in control cells (Fig. 2). The elimination of HRP from the
reaction mixture resulted in undetectable dityrosine formation,
indicating the requirement for peroxidative chemistry to mediate this
reaction and supporting the general reaction mechanism shown in Fig.
1a. In further support of this, the substitution of catalase
in place of HRP failed to generate any dityrosine (Fig. 2).
Additionally, cotreatment of catalase (1000 units/ml) with
HRP significantly inhibited TGF-
1-induced dityrosine formation (Fig.
2). Relatively high concentrations of catalase were required for
inhibition, most likely related to the fact that catalase has to
"compete" with HRP for H2O2, and the rate
constants for these reactions are several orders of magnitude higher
for HRP than for catalase (32). Almost complete inhibition of
TGF-
1-induced dityrosine formation was observed when cells were
preincubated with diphenyliodonium (DPI, 10 µM, added 15 min prior to measurement), a flavoprotein inhibitor that blocks
TGF-
1-induced H2O2 production (14) (Fig. 2).
Taken together, these results indicate the ability of
H2O2 generated by TGF-
1-stimulated fibroblasts to mediate HRP-catalyzed dimerization reactions involving the phenolic amino acid L-tyrosine in the extracellular
milieu of cultured fibroblasts.
1-induced
H2O2 is capable of inducing dimerization of
exogenous L-tyrosine, we hypothesized that such reactions
might occur on tyrosine residues of endogenous cellular or
extracellular proteins. We utilized a novel approach to test this
hypothesis by making use of a fluorophore-labeled phenolic compound
(tyramide-FITC) that undergoes similar reactions to those described for
p-substituted phenolic compounds (Fig. 1a;
see Fig. 1b for structural comparisons of tyrosine and
tyramide). In fact, this property of tyramide is the basis for its
utilization in signal amplification (31). This reagent was added to the same HBSS-based assay medium used in the L-tyrosine
experiments described above. The amplification diluent provided with
the assay kits for signal amplification was not used (because it
contains H2O2; see "Experimental
Procedures" for details). Fig. 3
demonstrates the characteristic fluorescent pattern observed when
TGF-
1-treated cells are incubated with tyramide-FITC and HRP.
Similar to TGF-
1-induced dityrosine formation, the elimination of
HRP from the reaction resulted in a loss of the fluorescent signal
(Fig. 3). Both catalase and DPI significantly inhibited the fluorescent
signal generated by the tyramide-FITC/HRP system (Fig. 3), suggesting a
requirement for H2O2 to mediate this effect.
These results suggest that TGF-
1-stimulated H2O2, in the presence of a heme peroxidase, is
capable of mediating cross-linking reactions between tyramide-FITC and
tyrosine residues of fibroblast or fibroblast-derived proteins. Based
on the localization of this fluorescent labeling reaction and pattern
of distribution, the protein(s) that are "targeted" by this
reaction seem to reside primarily in the ECM.
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Fig. 3.
Localization of endogenous proteins targeted
by H2O2/HRP-dependent tyrosine
cross-linking in cultured human lung fibroblasts. Control and
TGF- 1-treated (2 ng/ml × 16 h) cells were incubated in
assay medium containing fluorescein-labeled tyramide in the
absence/presence of HRP (5 units/ml) for 1 h followed by washing
and fluorescent microscopy (see "Experimental Procedures" and text
for details). The effect of catalase (1000 units/ml, coincubated during
the assay period) and the flavoprotein inhibitor, diphenyliodonium
(DPI, 10 µM added 15 min prior to assay) on
the TGF-
1-induced labeling (cross-linking) reaction is shown.
1) that do not
generate extracellular H2O2 on tyrosine
cross-linking utilizing the same tyramide-FITC assay system was
examined. Cells were exposed to varying concentrations of
H2O2 in the assay medium containing HRP and
tyramide-FITC for 5 min (see "Experimental Procedures" for
details). Fig. 4 demonstrates a
dose-dependent effect of H2O2 on
fluorescent labeling intensity. The pattern of fluorescent labeling was
similar to that observed with endogenous H2O2
production in TGF-
1-treated cells, suggesting that the same ECM
proteins may be targeted by these cross-linking reactions.
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Fig. 4.
Effect of exogenous
H2O2 on fluorophore-labeled
tyramide/HRP-dependent fluorescent intensity and pattern in
cultured lung fibroblasts. Cultured cells were incubated with
assay medium containing tyramide and HRP (5 units/ml) for 5 min. Cells
then were washed and viewed under a fluorescence microscope (see
"Experimental Procedures" for details). The effect of increasing
H2O2 concentrations (0, 1, 5, and 10 µM) on the intensity and pattern of fluorescent labeling
is shown.
DISCUSSION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
1-stimulated
fibroblasts are capable of mediating dimerization of
L-tyrosine by a peroxidative mechanism. Moreover, this
reaction seems to target tyrosine residues on endogenous protein
substrates, suggesting that this mechanism might be important for
mediating oxidative cross-linking of proteins under certain physiologic/pathologic conditions. This is the first report, to our knowledge, that demonstrates oxidative cross-linking reactions in
response to cytokine-generated ROS.
Interestingly, the ability of H2O2 to mediate
such reactions on tyrosine residues of ECM proteins has been well
demonstrated in the so-called oxidative burst of fertilization to
protect the freshly fertilized sea urchin oocyte (27) and to prevent
pathogen spread in the "plant hypersensitivity response" (34). Such
examples of a physiologic "defense mechanism" in humans are
lacking. The best, and likely only, known example of a "purposeful"
role for H2O2 in humans of heme
peroxidase-catalyzed dimerization reactions involving tyrosine is in
the biosynthesis of thyroxine, which is catalyzed by thyroid peroxidase
(35, 36). Under pathological conditions, cross-linking of
proteins by phagocyte-derived myeloperoxidase has been proposed as a
marker of oxidative stress in human atherosclerotic lesions (30).
1 and myofibroblasts in lesion formation and the involvement of phagocytic cells in this process provide all of the necessary components for this
reaction to occur in vivo. The recent data suggesting that
the biomechanical properties of the ECM may regulate myofibroblast differentiation are intriguing (15, 16). Oxidative cross-linking of ECM
would be expected to reduce tissue compliance and further promote
myofibroblast differentiation by this mechanism, suggesting a potential
positive feedback loop. Alternatively, cross-linking of ECM proteins
may render them less susceptible to digestion by matrix
metalloproteinases (39). If such effects can be confirmed in
vivo, it would present multiple potential targets for therapy of
fibrotic diseases including carefully designed antioxidant strategies.
1 in our study are currently unknown. It is
quite clear, however, that these proteins are localized primarily in
the ECM based not only on the observed pattern of fluorescent labeling
but also on the finding that the extracellular enzymes, HRP and
catalase, which do not freely cross the plasma membrane, regulate/alter
these reactions when added extracellularly. Candidate molecules for
this reaction include collagen and elastin, both of which have been
shown to undergo dityrosine-dependent cross-linking when
H2O2 and HRP are introduced exogenously (40, 41). Moreover, both of these ECM proteins are well recognized to be
up-regulated by TGF-
1 (42, 43). The possibility that such reactions
also may involve plasma membrane proteins at the cell surface requires
further study.
-amino group
of lysine side chains in these ECM proteins to form inter- and
intrachain cross-links (44). Interestingly, a "by-product" of this
reaction is H2O2. However, no role for lysyl oxidase-generated H2O2 in protein
cross-linking has been demonstrated. We have excluded the possibility
that lysyl oxidase activity may be responsible for TGF-
1-induced
H2O2 production. The lysyl oxidase inhibitors,
-aminopropionitrile and ethylenediamine, had no effect on
TGF-
1-induced NADH oxidase activity/H2O2
production (20). Moreover, we have demonstrated that this enzyme
activity involves a flavoprotein (14), which is not required for lysyl oxidase activity (45). Recent data suggest that the enzyme responsible for this effect of TGF-
1 in fibroblasts is related closely to the
phagocytic NAD(P)H oxidase family of plasma membrane
oxidases.2 Our ability to
measure extracellular H2O2 without detectable O
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ACKNOWLEDGEMENT |
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We thank Dr. Jay W. Heinecke (Washington University School of Medicine, St. Louis) for helpful discussions and suggestions with regard to the measurement of dityrosine.
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FOOTNOTES |
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* This work was supported by National Institutes of Health Grants K08 HL-03552 and HL-42376.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Pulmonary and Critical
Care Division, New England Medical Center, 750 Washington St., NEMC
257, Boston, MA 02111. Tel.: 617-636-7608; Fax: 617-636-5953; E-mail:
vthannickal@lifespan.org.
Published, JBC Papers in Press, February 6, 2001, DOI 10.1074/jbc.M100426200
2 J. M. Larios and V. J. Thannickal, manuscript in preparation.
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ABBREVIATIONS |
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The abbreviations used are:
TGF-1, transforming growth factor-
1;
ECM, extracellular matrix;
ROS, reactive oxygen species;
HRP, horseradish peroxidase;
HBSS, Hanks'
balanced salt solution;
FITC, fluorescein isothiocyanate.
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REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1. | Elias, J. A., Freundlich, B., Kern, J. A., and Rosenbloom, J. (1990) Chest 97, 1439-1445[Abstract] |
2. | Ward, P. A., and Hunninghake, G. W. (1998) Am. J. Respir. Crit. Care Med. 157, S123-S129[Medline] [Order article via Infotrieve] |
3. | Hogaboam, C. M., Smith, R. E., and Kunkel, S. L. (1998) Proc. Assoc. Am. Physicians 110, 313-320[Medline] [Order article via Infotrieve] |
4. |
Border, W. A.,
and Noble, N. A.
(1994)
N. Engl. J. Med.
331,
1286-1292 |
5. | Border, W. A., and Noble, N. A. (1995) J. Clin. Invest. 96, 655-656[Medline] [Order article via Infotrieve] |
6. | Powell, D. W., Mifflin, R. C., Valentich, J. D., Crowe, S. E., Saada, J. I., and West, A. B. (1999) Am. J. Physiol. 277, C1-C9[Medline] [Order article via Infotrieve] |
7. | Gauldie, J., Sime, P. J., Xing, Z., Marr, B., and Tremblay, G. M. (1999) Curr. Top. Pathol. 93, 35-45[Medline] [Order article via Infotrieve] |
8. | Desmouliere, A., Geinoz, A., Gabbiani, F., and Gabbiani, G. (1993) J. Cell Biol. 122, 103-111[Abstract] |
9. | Kuhn, C., and McDonald, J. A. (1991) Am. J. Pathol. 138, 1257-1265[Abstract] |
10. | Phan, S. H. (1996) Kidney Int. Suppl. 54, S46-S48[Medline] [Order article via Infotrieve] |
11. |
Dalton, S. L.,
Scharf, E.,
Davey, G.,
and Assoian, R. K.
(1999)
J. Biol. Chem.
274,
30139-30145 |
12. |
Finlay, G. A.,
Thannickal, V. J.,
Fanburg, B. L.,
and Paulson, K. E.
(2000)
J. Biol. Chem.
275,
27650-27656 |
13. | Thannickal, V. J., Aldweib, K. D., Rajan, T., and Fanburg, B. L. (1998) Biochem. Biophys. Res. Commun. 251, 437-441[CrossRef][Medline] [Order article via Infotrieve] |
14. |
Thannickal, V. J.,
and Fanburg, B. L.
(1995)
J. Biol. Chem.
270,
30334-30338 |
15. |
Arora, P. D.,
Narani, N.,
and McCulloch, C. A.
(1999)
Am. J. Pathol.
154,
871-882 |
16. | Narani, N., Arora, P. D., Lew, A., Luo, L., Glogauer, M., Ganss, B., and McCulloch, C. A. (1999) Curr. Top. Pathol. 93, 47-60[Medline] [Order article via Infotrieve] |
17. | Galbraith, C. G., and Sheetz, M. P. (1998) Curr. Opin. Cell Biol. 10, 566-571[CrossRef][Medline] [Order article via Infotrieve] |
18. | Streuli, C. (1999) Curr. Opin. Cell Biol. 11, 634-640[CrossRef][Medline] [Order article via Infotrieve] |
19. |
Katz, B. Z.,
Zamir, E.,
Bershadsky, A.,
Kam, Z.,
Yamada, K. M.,
and Geiger, B.
(2000)
Mol. Biol. Cell
11,
1047-1060 |
20. |
Thannickal, V. J.,
Day, R. M.,
Klinz, S. G.,
Bastien, M. C.,
Larios, J. M.,
and Fanburg, B. L.
(2000)
FASEB J.
14,
1741-1748 |
21. | Sundaresan, M., Yu, Z. X., Ferrans, V. J., Irani, K., and Finkel, T. (1995) Science 270, 296-299[Abstract] |
22. |
Bae, Y. S.,
Kang, S. W.,
Seo, M. S.,
Baines, I. C.,
Tekle, E.,
Chock, P. B.,
and Rhee, S. G.
(1997)
J. Biol. Chem.
272,
217-221 |
23. |
Brar, S. S.,
Kennedy, T. P.,
Whorton, A. R.,
Murphy, T. M.,
Chitano, P.,
and Hoidal, J. R.
(1999)
J. Biol. Chem.
274,
20017-20026 |
24. |
Barrett, W. C.,
DeGnore, J. P.,
Keng, Y. F.,
Zhang, Z. Y.,
Yim, M. B.,
and Chock, P. B.
(1999)
J. Biol. Chem.
274,
34543-34546 |
25. |
Thannickal, V. J.,
Aldweib, K. D.,
and Fanburg, B. L.
(1998)
J. Biol. Chem.
273,
23611-23615 |
26. | Halliwell, B., and Gutteridge, J. M. C. (1989) Free Radicals in Biology and Medicine , 2nd Ed. , pp. 99-101, Oxford University Press, New York |
27. | Shapiro, B. M. (1991) Science 252, 533-536[Medline] [Order article via Infotrieve] |
28. | Bradley, D. J., Kjellbom, P., and Lamb, C. J. (1992) Cell 70, 21-30[Medline] [Order article via Infotrieve] |
29. | Heinecke, J. W., Li, W., Francis, G. A., and Goldstein, J. A. (1993) J. Clin. Invest. 91, 2866-2872[Medline] [Order article via Infotrieve] |
30. |
Heinecke, J. W.,
Li, W.,
Daehnke, H. L. d.,
and Goldstein, J. A.
(1993)
J. Biol. Chem.
268,
4069-4077 |
31. |
Speel, E. J.,
Hopman, A. H.,
and Komminoth, P.
(1999)
J. Histochem. Cytochem.
47,
281-288 |
32. | Saunders, B. C., Holmes-Siedle, A. G., and Stark, B. P. (1964) Peroxidase: The Properties and Uses of a Versitile Enzyme and of Some Related Catalysts , Butterworths, Washington, D. C. |
33. | Thannickal, V. J., and Fanburg, B. L. (2000) Am. J. Physiol. 279, L1005-L1028 |
34. | Levine, A., Tenhaken, R., Dixon, R., and Lamb, C. (1994) Cell 79, 583-593[Medline] [Order article via Infotrieve] |
35. | Gorin, Y., Leseney, A. M., Ohayon, R., Dupuy, C., Pommier, J., Virion, A., and Deme, D. (1997) Biochem. J. 321, 383-388[Medline] [Order article via Infotrieve] |
36. |
De Deken, X.,
Wang, D.,
Many, M. C.,
Costagliola, S.,
Libert, F.,
Vassart, G.,
Dumont, J. E.,
and Miot, F.
(2000)
J. Biol. Chem.
275,
23227-23233 |
37. | Cross, C. E., Halliwell, B., Borish, E. T., Pryor, W. A., Ames, B. N., Saul, R. L., McCord, J. M., and Harman, D. (1987) Ann. Intern. Med. 107, 526-545[Medline] [Order article via Infotrieve] |
38. | Halliwell, B., Gutteridge, J. M., and Cross, C. E. (1992) J. Lab. Clin. Med. 119, 598-620[Medline] [Order article via Infotrieve] |
39. | Bailey, A. J. (2000) Wound Repair Regen. 8, 5-12[CrossRef][Medline] [Order article via Infotrieve] |
40. | LaBella, F., Keeley, F., Vivian, S., and Thornhill, D. (1967) Biochem. Biophys. Res. Commun. 26, 748-753[Medline] [Order article via Infotrieve] |
41. | LaBella, F., Waykole, P., and Queen, G. (1968) Biochem. Biophys. Res. Commun. 30, 333-338[Medline] [Order article via Infotrieve] |
42. |
Lawrence, R.,
Hartmann, D. J.,
and Sonenshein, G. E.
(1994)
J. Biol. Chem.
269,
9603-9609 |
43. | Marigo, V., Volpin, D., Vitale, G., and Bressan, G. M. (1994) Biochem. Biophys. Res. Commun. 199, 1049-1056[CrossRef][Medline] [Order article via Infotrieve] |
44. | Smith-Mungo, L. I., and Kagan, H. M. (1998) Matrix Biol. 16, 387-398[CrossRef][Medline] [Order article via Infotrieve] |
45. | Wang, S. X., Mure, M., Medzihradszky, K. F., Burlingame, A. L., Brown, D. E., Dooley, D. M., Smith, A. J., Kagan, H. M., and Klinman, J. P. (1996) Science 273, 1078-1084[Abstract] |
46. | Vissers, M. C., Day, W. A., and Winterbourn, C. C. (1985) Blood 66, 161-166[Abstract] |
47. | Nathan, C. F. (1987) J. Clin. Invest. 80, 1550-1560[Medline] [Order article via Infotrieve] |
48. | Suh, Y. A., Arnold, R. S., Lassegue, B., Shi, J., Xu, X., Sorescu, D., Chung, A. B., Griendling, K. K., and Lambeth, J. D. (1999) Nature 401, 79-82[CrossRef][Medline] [Order article via Infotrieve] |
49. |
Geiszt, M.,
Kopp, J. B.,
Varnai, P.,
and Leto, T. L.
(2000)
Proc. Natl. Acad. Sci. U. S. A.
97,
8010-8014 |