© 1999 American Heart Association, Inc.
Molecular Medicine |
Vascular Matrix Metalloproteinase-2 Cleaves Big Endothelin-1 Yielding a Novel Vasoconstrictor
From the Perinatal Research Centre (C.F-P., S.T.D.), Departments of Obstetrics/Gynaecology and Physiology, University of Alberta, Edmonton, Alberta, Canada and the Department of Pharmacology (M.W.R), University of Alberta, Edmonton, Alberta, Canada.
Correspondence to Sandra T. Davidge, Perinatal Research Centre, 232 HMRC, Departments of Obstetrics/Gynaecology and Physiology, University of Alberta, Edmonton, Alberta T6G 2S2, Canada. E-mail sandra.davidge{at}ualberta.ca
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Abstract |
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Abstract—Matrix metalloproteinase-2 (MMP-2, gelatinase A) and its tissue inhibitor (TIMP-2) are mainly known for their roles in the (patho)physiological remodeling of the vasculature, angiogenesis, tissue repair, tumor invasion, inflammation, and atherosclerotic plaque rupture. A mechanism of action of MMP-2 is the proteolytic breakdown of specific extracellular matrix proteins. The amino acid sequences in interstitial collagen (Gly-Leu/Ile) and laminin-5 (Ala-Leu) that are cleaved by MMP-2 are homologous to a region (Gly32-Leu33) within human big endothelin-1[1 to 38] (big ET-1). Big ET-1 requires cleavage to an active form to produce vasoconstriction. We tested the hypothesis that vascular MMP-2 can cleave big ET-1, thus generating a vasoconstrictor peptide. In perfused rat mesenteric arteries with an intact endothelium, inhibition of vascular MMP-2 with TIMP-2 reduced (by 16.2±4.2%) the vasoconstrictor effects of big ET-1 (50 pmol). However, when the endothelium was mechanically removed, TIMP-2 abolished (>90%) the vasoconstriction of big ET-1, and this effect was mimicked by an anti–MMP-2 antibody. Incubation of big ET-1 with recombinant human MMP-2 resulted in the specific cleavage of the Gly32-Leu33 bond of big ET-1. Moreover, the resultant peptide ET-1[1 to 32] exerted greater vasoconstrictor effects than big ET-1. We conclude that vascular MMP-2 contributes to the vasoconstrictor effects of big ET-1 by cleaving big ET-1 to yield a novel and potent vasoconstrictor, ET-1[1 to 32]. These data implicate, for the first time, the endogenous MMP-2/TIMP-2 system in the regulation of vascular reactivity.
Key Words: metalloproteinase • endothelin • vasculature
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Matrix metalloproteinase-2 (MMP-2, gelatinase A, EC 3.4.24.24) belongs to a family of zinc-dependent proteinases secreted from a variety of cell types, including vascular endothelial and smooth muscle cells.1 MMP-2 is expressed in a latent form (pro-MMP-2, 72 kDa) that is activated by a membrane-type-MMP located at the plasma membrane. Activation involves the removal of an amino terminal propeptide (
10 kDa), which converts pro-MMP-2 into MMP-2 (62 kDa)
that displays full proteolytic activity.1 2 3 MMP-2 is expressed in various cells and tissues, including the vascular smooth muscle and the endothelium.1 3 4 It is mainly known for its capacity to digest specific extracellular matrix proteins, such as collagen (types I, IV, V, VII, and X), elastin, fibronectin, and laminin-5.1 2 3 5 6 Thus, basal MMP-2 activity is associated with physiological remodeling, tissue repair, and angiogenesis.1 Tissue inhibitors of MMPs (TIMPs, particularly TIMP-2) regulate MMP-2 activity.2 Imbalance in the expression of MMP-2/TIMP-2 leading to excessive matrix degradation by MMP-2 has been correlated with tissue injury, tumor invasion, inflammation, atherosclerosis, and atherosclerotic plaque rupture.1 2 3
Recently, a novel pathway of platelet aggregation was discovered that is mediated by the release of MMP-2 from activated platelets,7 which suggests that MMP-2 may have as-yet-unknown regulatory actions in the cardiovascular system. In seeking a role for MMP-2 in the regulation of vascular reactivity that is unrelated to long-term extracellular matrix breakdown, we investigated whether MMP-2 could mediate the activation of inactive precursors of vasoactive peptides/proteins, such as big endothelin-1[1–38] (big ET-1).7
Similar to MMP-2, big ET-1 is produced and secreted by the endothelium and the vascular smooth muscle.8 9 Big ET-1 is essentially inactive, and it requires activation to produce vasoconstriction. Serine and aspartic proteases, as well as metalloproteinases, can cleave big ET-1, yielding peptides with various vasoconstrictor potencies.8 9 10 11 12 13 14 15 16 17 18 In the cardiovascular system, zinc-dependent endoproteases, termed endothelin-converting enzymes (ECEs), may mediate the vasoconstrictor effects of big ET-1.10 12 17 ECEs cleave big ET-1 at the Val21-Trp22 bond, yielding the potent vasoconstrictor ET-1[1–21].8 10 11 12 18 In the airway, chymase from mast cells was recently shown to cleave big ET-1 at the Tyr31-Gly32 peptide bond, yielding ET-1[1–31], which is a vasoconstrictor of tracheal and vascular smooth muscle and could be involved in allergic inflammation.18 All these observations reveal different pathways of activation of big ET-1. It is likely that these pathways are differentially expressed, depending on the localization of the respective big ET-1-activating protease and the tissue targeted by the active form of big ET-1 that is generated. Interestingly, a region exists within human big ET-1 (Gly32-Leu33) that is homologous with the amino acid sequences in interstitial collagen (Gly-Leu/Ile)5 and laminin-5 (Ala-Leu)6 that are cleaved by MMP-2 during some (patho)physiological processes. Therefore, we hypothesized that vascular MMP-2 could specifically cleave big ET-1 at the Gly32-Leu33 bond, thus generating a vasoconstrictor peptide.
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Materials and Methods |
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Western Blot and Gelatin Zymography
Immunodetection and gelatin zymography of MMP-2 from rat mesenteric arteries were performed as described previously.1 7
Immunohistochemistry
Serial cryostat sections (8 µm) of mesenteric arteries
were deposited on glass slides and incubated for 60 minutes with either
anti–MMP-2 monoclonal antibody or control mouse IgG (Rose
Scientific) diluted in gelatinase-free fetal calf serum.
Development was performed with cyanine-conjugated anti-mouse IgG
(BIO/CAN Scientific). Sections were inspected for MMP-2
immunoreactivity using a fluorescence microscope (Olympus BX40,
Carsen Group Inc).
In Situ Zymography
Regions of the arterial wall with net MMP-2
proteolytic activity were localized by in situ
zymography19 as follows. The gelatin coating of a
negative Polaroid film served as an immobilized substrate
for in situ zymography. Each cryostat section of the mesenteric artery
was layered with 2 µg of highly (>95%) purified, solubilized
collagen type IV (Calbiochem). Then, each cryostat section was covered
with a piece (25 mm2) of Polaroid film,
with the gelatin-coated side of the film facing the cryostat section.
This setting was further topped with a coverslip and left in a
humidified chamber. After 48 hours, proteolytic activity was
recorded (Olympus BX40 microscope). As a control, some of the
sections were supplemented with TIMP-1 (2 µmol/L, Chemicon),
which inhibits both MMP-2 and MMP-9.19
Chemical and Enzymatic Reactions
To study the interactions between big ET-1 and MMP-2, the
general strategy was to incubate synthetic big ET-1 with highly pure
recombinant human MMP-2 (Chemicon International) for times ranging from
1 to 48 hours. The incubation products were analyzed by
high-performance liquid chromatography, mass
spectrometry, amino acid analysis, and bioassay. Incubations
were performed at 37°C in HEPES-phosphate saline solution (pH 7.4)
composed of the following (in mmol/L): NaCl 142, KCl 4.7,
MgSO4 1.17, CaCl2 1.56,
HEPES 10, and KH2PO4
1.18.
Microperfusion Bioassay
The effect of endothelin peptides on smooth muscle reactivity
was studied using rat mesenteric arteries. Animal protocols were
conducted in accordance with institutional guidelines issued by the
Canada Council on Animal Care. Male Sprague-Dawley rats (350 to
450 g) were anesthetized with methohexital sodium (50
mg/kg) and exsanguinated. Arteries (209±12 µm, inner diameter;
1 to 1.5 mm, length) were dissected from fat tissue and
adventitia. All experiments were conducted on cannulated arteries
perfused at a constant temperature (37°C) and flow rate (10 µL/min)
with HEPES-phosphate saline solution-glucose (5.5 mmol/L). In some
experiments, the arteries were denuded of endothelium.
Endothelium removal was performed mechanically using a
human hair threaded through the lumen of the artery and rubbed back and
forth.20 Small volumes (1 to 5 µL) of big ET-1,
ET-1[1–21], MMP-2 cleavage products of big ET-1, or specified
drugs were injected into the perfusion line toward the mesenteric
artery by using a high-performance liquid
chromatography injection valve (Rheodyne Model 9725I,
Mandel Scientific Co) provided with a 20-µL loop.
Statistics
Results are the mean±SEM of at least 3 independent experiments.
They were analyzed using 1-way ANOVA. When significant
differences were found, the Tukey multiple comparisons’ test was used
(Jandel SigmaStat statistical software). P<0.05 was
considered statistically significant.
An expanded Materials and Methods section is available online at http://www.circresaha.org.
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Results |
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Identification and Involvement of MMP-2 in the Vasoconstrictor Effects of Big ET-1 on Rat Mesenteric Arteries
MMP-2 was the dominant gelatinolytic metalloproteinase detected on the rat mesenteric arteries, as confirmed by gelatin-zymography and Western blotting (Figure 1A
). Net MMP-2 proteolytic activity was
detected on medial smooth muscle cells (Figure 1B), suggesting
that the MMP-2 activity localized therein was higher than the activity
of the TIMPs. Furthermore, MMP-2 immunoreactivity was predominant on
smooth muscle cells of the intima and media (Figure 1C).
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Big ET-1 (0.5 to 50 pmol), when infused through the arteries, resulted
in dose-dependent vasoconstriction. In arteries with an intact
endothelium, coinfusion of TIMP-2 (0.17 to 10 pmol)
with big ET-1 (50 pmol) partially inhibited the vasoconstrictor effects
of big ET-1, with a maximum effect of 16.2±4.2% (P<0.05;
n=3). In contrast, in endothelium-denuded arteries,
coinfusion of TIMP-2 abolished the vasoconstrictor effects of big ET-1
(Figure 2). TIMP-2 exerted no significant
effect on the vasoconstrictor response to ET-1[1–21] (Figure 2). Moreover, an MMP-2–specific antibody, but not
control IgG, inhibited the vasoconstrictor effects of big ET-1 (Figure 3). These results showed that vascular
MMP-2 is, indeed, involved in the vasoconstrictor effects of big ET-1.
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MMP-2), but not control rabbit IgG, dose-dependently inhibited
vasoconstriction of big ET-1 (50 pmol). Results are
representative of 3 separate experiments.
MMP-2 Cleaves Big ET-1, Yielding Novel Peptides
We next investigated whether MMP-2 could cleave big ET-1 and
generate a bioactive peptide. Big ET-1 was incubated with recombinant
human pro-MMP-2 (6% of its gelatinolytic activity
was due to MMP-2, as determined by gelatin-zymography). This resulted
in partial conversion of big ET-1 into 2 fragments (X and Y; Figure 4, A through C), that were well resolved
chromatographically from big ET-1 and ET-1[1–21]. When
big ET-1 was incubated with activated MMP-2, >90% of big ET-1
was converted into X and Y fragments (Figure 4D). However, MMP-2
did not cleave ET-1[1–21] (data not shown). MMP-2–dependent
conversion of big ET-1 was inhibited in the presence of TIMP-2 (Figure 4E). No further peptides of big ET-1 were generated when big
ET-1 and MMP-2 were incubated for 
48 hours, suggesting the presence
of a single cleavage site on big ET-1. Microsequence and mass
spectrometry analyses identified the peptides X and Y as the
fragments Leu33-Ser38 and
Cys1-Gly32 of big ET-1,
respectively, confirming that the cleavage site on big ET-1 was the
MMP-2–susceptible
Gly32-Leu33 bond
(Table). Cleavage at
Gly32-Leu33 was also
observed when human big ET-1 was replaced by rat big
ET-1[1–39], reinforcing the specificity of MMP-2 for this
peptide bond. Peptide Y (3685 Da) was larger than ET-1[1–21] (2492
Da) but smaller than big ET-1 (4284 Da), and it was termed
ET-1[1–32]. ET-1[1–32], but not peptide X
(Leu33-Ser38), comprised
the full sequence of ET-1[1–21], thus containing the binding domain
to endothelin receptors.
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ET-1[1–32] Is a Novel Vasoconstrictor
ET-1[1–32] induced dose-dependent vasoconstriction of the
mesenteric arteries (Figure 5) that
lasted >20 to 30 minutes after its infusion (Figure 6). Furthermore, the vasoconstrictor
effects of ET-1[1–32] were significantly (P<0.05)
greater than those of its precursors, big ET-1 and ET-1[1–21], such
that ET-1[1–32] ED50=4.0±1.1 pmol
<ET-1[1–21] ED50=27.7±8.2 pmol <big ET-1
ED50=45.7±5.1 pmol (Figure 4
).
Preinfusion of the selective ETA receptor
antagonist N-acetyl-[D-Trp16]-ET-1
(20 nmol) before the infusion of ET-1[1–32] (2 pmol)
inhibited vasoconstriction to this peptide by 15±1% (n=3).
Preinfusion of the artery with PD 142893 (20 nmol), a
nonselective antagonist of ETA and
ETB receptors,21 abolished the
vasoconstrictor effects of ET-1[1–32] but not those of
phenylephrine (Figure 6).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We investigated the impact of vascular MMPs on the vasoconstrictor effects of big ET-1 on rat mesenteric arteries. Zymography and Western blot analysis revealed that MMP-2 was the major gelatinase expressed in these arteries. In situ zymography showed that the net gelatinolytic activity in the arteries colocalized with immunoreactive MMP-2. To study the significance of vascular MMP-2 for the vasoconstrictor effects of big ET-1, TIMP-2 (an endogenous inhibitor of MMPs with preferred effects on MMP-21 ) or an MMP-2–specific antibody were administered together with big ET-1. The selective inhibition of MMP-2 significantly reduced the vasoconstrictor effects of big ET-1 but not ET-1[1–21]. These data clearly indicate that vascular MMP-2 contributed to the vasoconstrictor effects of big ET-1 on arteries.
To investigate the mechanism of this action of MMP-2, we incubated big ET-1 with recombinant human MMP-2. The reaction resulted in the specific cleavage of big ET-1 at the Gly32-Leu33 bond, yielding a novel vasoconstrictor peptide, ET-1[1–32]. A comparison of the vasoconstrictor effects of ET-1[1–32] with those of big ET-1 and ET-1[1–21] showed that ET-1[1–32] was the most potent vasoconstrictor on rat mesenteric arteries. Moreover, ETA and ETB receptors are likely to mediate the vasoconstrictor effects of ET-1[1–32], because the blockade of both ETA and ETB receptors is required to abolish the vasoconstrictor effects of this peptide. It is, however, unlikely that a conversion of ET-1[1–32] into ET-1[1–21] by vascular ECEs is obligatory for expression of vasoactivity, because ET-1[1–32] lacks the COOH-terminus of big ET-1, which is necessary for recognition and cleavage by ECE.10 Hence, ET-1[1–32] may represent a new vasoconstrictor form of big ET-1 that is generated after the activation of MMP-2.
Pharmacological studies of TIMP-2 and MMP-2 antibody showed that the interaction between big ET-1 and vascular MMP-2 was more prominent in the mesenteric arteries denuded of endothelium than in those with an intact endothelium. These observations are of high biological significance. In contrast to ECE, which is expressed mainly by the vascular endothelium,10 the net proteolytic activity and immunoreactivity of MMP-2 was more pronounced in the intimal and medial vascular smooth muscle cells of the arteries studied. These results agreed with recent reports showing that MMP-2 concentrates along the basement membrane and on those smooth muscle cells in close proximity to the intima.1 2 3 4 Thus, an activation of big ET-1 to ET-1[1–21] may be favored in the endothelium, where ECE activity is high. In contrast, conversion of big ET-1 into ET-1[1–32] by MMP-2 may be favored in vascular smooth muscle cells, where the activity of ECE is lower.
Interestingly, concerted overexpression of MMP-2 and big ET-1 can be observed at sites of tissue remodeling and repair under conditions of tissue injury, inflammation, and cancer. Moreover, vascular pathologies such as hypertension, atherosclerosis, atherosclerotic plaque rupture, and restenosis are also associated with the increased expression of MMPs and endothelins.1 22 23 24 25 26 27 28 29 Endothelial injury and increased generation of these mediators in the vessel wall–platelet microenvironment are likely to promote vasospasm, platelet activation, smooth-muscle-cell migration, proliferation, and exaggerated expansion of the extracellular matrix.7 28 29 In line with this, recent results from our laboratory suggest that the generation of ET-1[1–32] occurred in conditions associated with the generation of thrombin, a key enzyme in the response to tissue injury (C. Fernandez-Patron, M.W. Radomski, and S.T. Davidge, unpublished data, 1999). Indeed, we found that a rapid release of bioactive MMP-2 occurred in response to thrombin-stimulation in rat mesenteric and aorta arteries.30 Furthermore, the vasoactive effects of thrombin were inhibited by TIMP-2 and abolished by endothelin receptor antagonists. Thus, we propose that an activation of big ET-1 to ET-1[1–32] by vascular MMP-2 may contribute to the vasoactive effects of thrombin.
The discovery of the MMP-2–dependent pathway for the generation of ET-1[1–32] may have important pharmacological significance. A large effort has been devoted to the development of synthetic selective inhibitors of MMPs.1 These compounds are now being clinically tested for their ability to ameliorate the course of inflammatory and cancer diseases.1 Therefore, selective inhibition of MMP-2 may represent a new pharmacological strategy for regulating vascular reactivity in pathological conditions in which MMP-2 and big ET-1 are overexpressed.
We have not identified a significant expression of gelatinolytic MMPs other than MMP-2 in the rat mesenteric arteries. However, MMPs such as MMP-1 and MMP-9 share many substrates with MMP-21 and may also cleave big ET-1. In addition, an increasing number of different pathways of activation of big ET-1 exist (eg, via ECE, mast cell chymase, and MMP-2) leading to different bioactive ET-1 peptides (eg, ET-1[1–21], ET-1[1–31], and ET-1[1–32]). Further studies are needed to clarify the contribution of ET-1[1–32] relative to ET-1[1–21] and ET-1[1–31] in vascular biology and disease. It is likely that these peptides play different roles in (patho)physiology, depending on the tissue expression of the respective big ET-1-activating proteases.
In summary, we found that vascular MMP-2 contributed to the vasoconstrictor effects of big ET-1 on rat mesenteric arteries via the generation of a novel vasoconstrictor peptide, ET-1[1–32]. This pathway may have important significance for the physiology, pathology, and pharmacology of the vessel wall.
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Acknowledgments |
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This work was supported by grants from the Medical Research Council of Canada (MRC; MT-13404 [and MT-14074 to M.W. Radomski]) and the Heart and Stroke Foundation of Canada (to S.T. Davidge). C. Fernandez-Patron is a postdoctoral fellow of the Alberta Heritage Foundation for Medical Research (AHFMR), the Canadian Hypertension Society, and the Medical Research Council of Canada. S.T. Davidge is a scholar of AHFMR and the Heart and Stroke Foundation of Canada. M.W. Radomski is a scholar of AHFMR.
Received June 8, 1999; accepted September 15, 1999.
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References |
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|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Parks WC, Mecham RP, eds. Matrix Metalloproteinases. San Diego, Calif: Academic Press; 1998.
2. Ray JM, Stetler-Stevenson WG. The role of metalloproteinases and their inhibitors in tumor, metastasis, and angiogenesis. Eur Respir J. 1994;7:2062–2072.[Abstract]
3. Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opinion Cell Biol. 1995;7:728–735.[Medline] [Order article via Infotrieve]
4.
Li Z, Froehlich J, Galis ZS, Lakatta EG. Increased
expression of matrix metalloproteinase-2 in the thickened intima of
aged rats. Hypertension. 1999;33:116–123.
5.
Aimes RT, Quigley JP. Matrix metalloproteinase-2 is an
interstitial collagenase:
inhibitor-free enzyme catalyzes the cleavage of type I
collagen generating the specific 3/4- and 1/4-length
fragments. J Biol Chem. 1995;270:5872–5876.
6.
Gianelli G, Falk-Marzillier J, Shiraldi O,
Stetler-Stevenson W, Quaranta V. Induction of cell migration by matrix
metalloprotease-2 cleavage of laminin-5. Science. 1997;277:225–228.
7. Sawicki G, Salas E, Murat J, Miszta-Lane H, Radomski MW. Release of gelatinase A during platelet activation mediates aggregation. Nature. 1997;386:616–619.[Medline] [Order article via Infotrieve]
8. Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988;332:411–415.[Medline] [Order article via Infotrieve]
9. Yu JC, Davenport AP. Secretion of endothelin-1 and endothelin-3 by human cultured vascular smooth muscle cells. Br J Pharmacol. 1995;114:551–557.[Medline] [Order article via Infotrieve]
10. Xu D, Emoto N, Giaid A, Slaughter C, Kaw S, de Wit D, Yanagisawa M. ECE-1: a membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1. Cell. 1994;78:473–485.[Medline] [Order article via Infotrieve]
11. Opgenorth TJ, Wu-Wong JR, Shiosaki K. Endothelin-converting enzymes. FASEB J. 1992;6:2653–2659.[Abstract]
12.
McMahon EG, Palomo MA, Moore WM, McDonald JF, Stern MK.
Phosphoramidon blocks the pressor activity of porcine
big endothelin-1 (1–39) in vivo and conversion of big endothelin-1
(1–39) to endothelin-1 (1–21) in vitro. Proc Natl Acad Sci
U S A. 1991;88:703–707.
13. Kimura S, Kasuya Y, Sawamura T, Osamu S, Yoshiki S, Yanagisawa M, Goto K, Masaki T. Conversion of big endothelin-1 to 21-residue endothelin-1 is essential for expression of full vasoconstrictor activity: structure-activity relationships of big endothelin-1. J Cardiovasc Pharmacol. 1989;13(suppl 5):S5–S7.
14. Patterson K, MacNaul R, Rubanyi GM, Parker-Botello LH. Separation and biological activity of the chymotrypsin and cathepsin G cleavage products of big endothelin (1–39). FASEB J. 1990;4:A909. Abstract.
15.
Kaw S, Hecker M, Vane JR. The two step conversion of
big endothelin 1 to endothelin 1 by subcellular fractions from human
polymorphonuclear leukocytes. Proc Natl Acad Sci
U S A. 1992;89:6886–6890.
16. Ahn K, Sisneros AM, Herman SB, Pan SM, Hupe D, Lee C, Nikam S, Cheng X, Doherty AM, Schroeder RL, Haleen SJ, Kaw S, Emoto N, Yanagisawa M. Novel selective quinazoline inhibitors of endothelin converting enzyme-1. Biochem Biophys Res Commun. 1998;243:184–190.[Medline] [Order article via Infotrieve]
17.
Emoto N, Yanagisawa M. Endothelin-converting enzyme-2
is a membrane-bound, phosphoramidon sensitive
metalloprotease with acidic pH optimum. J Biol Chem. 1995;270:15262–15268.
18. Kishi F, Minami K, Okishima N, Murakami M, Mori S, Yano M, Niwa Y, Nakaya Y, Kido H. Novel 31-amino-acid-length endothelins cause constriction of vascular smooth muscle. Biochem Biophys Res Commun. 1998;248:387–390.[Medline] [Order article via Infotrieve]
19. Galis ZS, Sukhova GK, Libby P. Microscopic localization of active proteases by in situ zymography: detection of matrix metalloproteinase activity in vascular tissue. FASEB J. 1995;9:974–980.[Abstract]
20. Osol GC, Cipolla M, Knutson S. A new method for mechanically denuding the endothelium of small (50–150 µm) arteries with a human hair. Blood Vessels. 1989;26:320–324.[Medline] [Order article via Infotrieve]
21. Hingorani G, Major T, Panek R, Flynn M, Reynolds E, He X, Cody W, Doherty A, Rapundalo S. In vitro pharmacology of a non-selective (ETA/ETB) endothelin receptor antagonist, PD 142893 (Ac-(ß-phenyl)D-Phe-L-Leu-L-Asp-L-Ile-Ile-L-Trp trifluoroacetate). FASEB J. 1992;6:A1003. Abstract.
22. Battistini B, Chailler P, D’Orleans-Juste P, Briere N, Sirois P. Growth regulatory properties of endothelins. Peptides. 1993;14:385–399.[Medline] [Order article via Infotrieve]
23. Rubanyi GM, Polokoff MA. Endothelins: molecular biology, biochemistry, pharmacology, physiology, and pathophysiology. Pharmacol Rev. 1994;46:325–415.[Medline] [Order article via Infotrieve]
24. Ishibashi M, Ito N, Fujita M, Furue H, Yamaji T. Endothelin-1 as an aggravating factor of diseminated intravascular coagulation associated with malignant neoplasms. Cancer. 1994;73:191–195.[Medline] [Order article via Infotrieve]
25.
Jenkins GM, Crow MT, Bilato C, Gluzband Y, Ryu WS, Li
Z, Stetler-Stevenson W, Nater C, Froehlich JP, Lakatta EG, Cheng L.
Increased expression of membrane-type matrix metalloproteinase and
preferential localization of matrix metalloproteinase-2 to the
neointima of balloon-injured rat carotid arteries.
Circulation. 1998;97:82–90.
26. Newby AC, Southgate KM, Davies M. Extracellular matrix degrading metalloproteinases in the pathogenesis of arteriosclerosis. Basic Res Cardiol. 1993;89:59–70.
27. Libby P, Geng Y-J, Sukhova GK, Simon DI, Lee RT, Alexander RW, Clowes A, Tanaka K, Fuster V, Kume N, Suzuki A. Molecular determinants of atherosclerotic plaque vulnerability. Ann N Y Acad Sci. 1997;811:134–145.[Medline] [Order article via Infotrieve]
28.
Kirchegast M, Muenter K. Endothelins and
restenosis. Cardiovasc Res. 1998;39:550–555.
29. Libby P, Tanaka H. The molecular basis of restenosis. Prog Cardiovasc Dis.. 1997;40:97–106.[Medline] [Order article via Infotrieve]
30. Fernandez-Patron C, Zhang Y, Radomski MW, Hollenberg MD, Davidge ST. Rapid release of matrix metalloproteinase (MMP)-2 by thrombin in the rat aorta: modulation by protein tyrosine kinase/phosphatase. Thromb Haemost. 1999;82:1353–1357.[Medline] [Order article via Infotrieve]
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 K. J. Greenlee, Z. Werb, and F. Kheradmand Matrix Metalloproteinases in Lung: Multiple, Multifarious, and Multifaceted Physiol Rev, January 1, 2007; 87(1): 69 - 98. [Abstract] [Full Text] [PDF]
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 J. L. Wright, H. Tai, R. Wang, X. Wang, and A. Churg Cigarette smoke upregulates pulmonary vascular matrix metalloproteinases via TNF-{alpha} signaling Am J Physiol Lung Cell Mol Physiol, January 1, 2007; 292(1): L125 - L133. [Abstract] [Full Text] [PDF]
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J. Z. He, A. Quan, Y. Xu, H. Teoh, G. Wang, J. E. Fish, B. M. Steer, S. Itohara, P. A. Marsden, S. T. Davidge, et al. Induction of matrix metalloproteinase-2 enhances systemic arterial contraction after hypoxia Am J Physiol Heart Circ Physiol, January 1, 2007; 292(1): H684 - H693. [Abstract] [Full Text] [PDF]
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 J. Verstappen and J.W. Von den Hoff Tissue Inhibitors of Metalloproteinases (TIMPs): Their Biological Functions and Involvement in Oral Disease Journal of Dental Research, December 1, 2006; 85(12): 1074 - 1084. [Abstract] [Full Text] [PDF]
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C. Kusmic, G. Lazzerini, F. Coceani, R. Barsacchi, A. L'Abbate, and G. Sambuceti Paradoxical coronary microcirculatory constriction during ischemia: a synergic function for nitric oxide and endothelin Am J Physiol Heart Circ Physiol, October 1, 2006; 291(4): H1814 - H1821. [Abstract] [Full Text] [PDF]
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 J. Herrmann, L. O. Lerman, D. Mukhopadhyay, C. Napoli, and A. Lerman Angiogenesis in Atherogenesis Arterioscler Thromb Vasc Biol, September 1, 2006; 26(9): 1948 - 1957. [Abstract] [Full Text] [PDF]
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 E. Thomson, P. Kumarathasan, and R. Vincent Pulmonary Expression of PreproET-1 and PreproET-3 mRNAs Is Altered Reciprocally in Rats After Inhalation of Air Pollutants. Experimental Biology and Medicine, June 1, 2006; 231(6): 979 - 984. [Abstract] [Full Text] [PDF]
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 A. Jeyabalan, L. J. Kerchner, M. C. Fisher, J. T. McGuane, K. D. Doty, and K. P. Conrad Matrix metalloproteinase-2 activity, protein, mRNA, and tissue inhibitors in small arteries from pregnant and relaxin-treated nonpregnant rats J Appl Physiol, June 1, 2006; 100(6): 1955 - 1963. [Abstract] [Full Text] [PDF]
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 C. Franco, B. Ho, D. Mulholland, G. Hou, M. Islam, K. Donaldson, and M. P. Bendeck Doxycycline Alters Vascular Smooth Muscle Cell Adhesion, Migration, and Reorganization of Fibrillar Collagen Matrices Am. J. Pathol., May 1, 2006; 168(5): 1697 - 1709. [Abstract] [Full Text] [PDF]
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H. Nagase, R. Visse, and G. Murphy Structure and function of matrix metalloproteinases and TIMPs Cardiovasc Res, February 15, 2006; 69(3): 562 - 573. [Abstract] [Full Text] [PDF]
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G.-Y. Wang, M. R. Bergman, A. P. Nguyen, S. Turcato, P. M. Swigart, M. C. Rodrigo, P. C. Simpson, J. S. Karliner, D. H. Lovett, and A. J. Baker Cardiac transgenic matrix metalloproteinase-2 expression directly induces impaired contractility Cardiovasc Res, February 15, 2006; 69(3): 688 - 696. [Abstract] [Full Text] [PDF]
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 D. C. Souza-Costa, T. Zerbini, A. C. Palei, R. F. Gerlach, and J. E. Tanus-Santos L-arginine Attenuates Acute Pulmonary Embolism-Induced Increases in Lung Matrix Metalloproteinase-2 and Matrix Metalloproteinase-9 Chest, November 1, 2005; 128(5): 3705 - 3710. [Abstract] [Full Text] [PDF]
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 E. Thomson, P. Kumarathasan, P. Goegan, R. A. Aubin, and R. Vincent Differential Regulation of the Lung Endothelin System by Urban Particulate Matter and Ozone Toxicol. Sci., November 1, 2005; 88(1): 103 - 113. [Abstract] [Full Text] [PDF]
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 A. C.T. Palei, R. A.G. Zaneti, G. M. Fortuna, R. F. Gerlach, and J. E. Tanus-Santos Hemodynamic Benefits of Matrix Metalloproteinase-9 Inhibition by Doxycycline During Experimental Acute Pulmonary Embolism Angiology, September 1, 2005; 56(5): 611 - 617. [Abstract] [PDF]
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J. Chen, C.-H. Tung, J. R. Allport, S. Chen, R. Weissleder, and P. L. Huang Near-Infrared Fluorescent Imaging of Matrix Metalloproteinase Activity After Myocardial Infarction Circulation, April 12, 2005; 111(14): 1800 - 1805. [Abstract] [Full Text] [PDF]
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D. O. Debrah, K. P. Conrad, L. A. Danielson, and S. G. Shroff Effects of relaxin on systemic arterial hemodynamics and mechanical properties in conscious rats: sex dependency and dose response J Appl Physiol, March 1, 2005; 98(3): 1013 - 1020. [Abstract] [Full Text] [PDF]
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 M. M. Lalu, E. Pasini, C. J. Schulze, M. Ferrari-Vivaldi, G. Ferrari-Vivaldi, T. Bachetti, and R. Schulz Ischaemia-reperfusion injury activates matrix metalloproteinases in the human heart Eur. Heart J., January 1, 2005; 26(1): 27 - 35. [Abstract] [Full Text] [PDF]
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 K. P. Conrad Mechanisms of Renal Vasodilation and Hyperfiltration During Pregnancy Reproductive Sciences, October 1, 2004; 11(7): 438 - 448. [Abstract] [PDF]
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 K. P. Conrad and J. Novak Emerging role of relaxin in renal and cardiovascular function Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2004; 287(2): R250 - R261. [Abstract] [Full Text] [PDF]
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 B.A. Kelly, B.C. Bond, and L. Poston Aortic adaptation to pregnancy: elevated expression of matrix metalloproteinases-2 and -3 in rat gestation Mol. Hum. Reprod., May 1, 2004; 10(5): 331 - 337. [Abstract] [Full Text] [PDF]
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 G G Koliakos, A G P Konstas, U Schlotzer-Schrehardt, G Hollo, D Mitova, D Kovatchev, S Maloutas, and N Georgiadis Endothelin-1 concentration is increased in the aqueous humour of patients with exfoliation syndrome Br J Ophthalmol, April 1, 2004; 88(4): 523 - 527. [Abstract] [Full Text] [PDF]
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 I. A. Arenas, Y. Xu, P. Lopez-Jaramillo, and S. T. Davidge Angiotensin II-induced MMP-2 release from endothelial cells is mediated by TNF-{alpha} Am J Physiol Cell Physiol, April 1, 2004; 286(4): C779 - C784. [Abstract] [Full Text] [PDF]
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K. Conant, C. St. Hillaire, H. Nagase, R. Visse, D. Gary, N. Haughey, C. Anderson, J. Turchan, and A. Nath Matrix Metalloproteinase 1 Interacts with Neuronal Integrins and Stimulates Dephosphorylation of Akt J. Biol. Chem., February 27, 2004; 279(9): 8056 - 8062. [Abstract] [Full Text] [PDF]
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 K. Kohlstedt, R. P. Brandes, W. Muller-Esterl, R. Busse, and I. Fleming Angiotensin-Converting Enzyme Is Involved in Outside-In Signaling in Endothelial Cells Circ. Res., January 9, 2004; 94(1): 60 - 67. [Abstract] [Full Text] [PDF]
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L. Hao, M. Du, A. Lopez-Campistrous, and C. Fernandez-Patron Agonist-Induced Activation of Matrix Metalloproteinase-7 Promotes Vasoconstriction Through the Epidermal Growth Factor-Receptor Pathway Circ. Res., January 9, 2004; 94(1): 68 - 76. [Abstract] [Full Text] [PDF]
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A. Jeyabalan, J. Novak, L. A. Danielson, L. J. Kerchner, S. L. Opett, and K. P. Conrad Essential Role for Vascular Gelatinase Activity in Relaxin-Induced Renal Vasodilation, Hyperfiltration, and Reduced Myogenic Reactivity of Small Arteries Circ. Res., December 12, 2003; 93(12): 1249 - 1257. [Abstract] [Full Text] [PDF]
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C. J. Schulze, W. Wang, W. L. Suarez-Pinzon, J. Sawicka, G. Sawicki, and R. Schulz Imbalance Between Tissue Inhibitor of Metalloproteinase-4 and Matrix Metalloproteinases During Acute Myoctardial Ischemia-Reperfusion Injury Circulation, May 20, 2003; 107(19): 2487 - 2492. [Abstract] [Full Text] [PDF]
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Y. Xu, I. A Arenas, S. J Armstrong, and S. T Davidge Estrogen modulation of left ventricular remodeling in the aged heart Cardiovasc Res, February 1, 2003; 57(2): 388 - 394. [Abstract] [Full Text] [PDF]
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C. Qun Gao, G. Sawicki, W. L Suarez-Pinzon, T. Csont, M. Wozniak, P. Ferdinandy, and R. Schulz Matrix metalloproteinase-2 mediates cytokine-induced myocardial contractile dysfunction Cardiovasc Res, February 1, 2003; 57(2): 426 - 433. [Abstract] [Full Text] [PDF]
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W. Wang, C. J. Schulze, W. L. Suarez-Pinzon, J. R.B. Dyck, G. Sawicki, and R. Schulz Intracellular Action of Matrix Metalloproteinase-2 Accounts for Acute Myocardial Ischemia and Reperfusion Injury Circulation, September 17, 2002; 106(12): 1543 - 1549. [Abstract] [Full Text] [PDF]
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P. Jurasz, A. W.Y. Chung, A. Radomski, and M. W. Radomski Nonremodeling Properties of Matrix Metalloproteinases: The Platelet Connection Circ. Res., May 31, 2002; 90(10): 1041 - 1043. [Full Text] [PDF]
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Z. S. Galis and J. J. Khatri Matrix Metalloproteinases in Vascular Remodeling and Atherogenesis: The Good, the Bad, and the Ugly Circ. Res., February 22, 2002; 90(3): 251 - 262. [Abstract] [Full Text] [PDF]
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W. Wang, G. Sawicki, and R. Schulz Peroxynitrite-induced myocardial injury is mediated through matrix metalloproteinase-2 Cardiovasc Res, January 1, 2002; 53(1): 165 - 174. [Abstract] [Full Text] [PDF]
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 N. Levy, M. Gordin, R. Mamluk, M. Yanagisawa, M. F. Smith, J. H. Hampton, and R. Meidan Distinct Cellular Localization and Regulation of Endothelin-1 and Endothelin-Converting Enzyme-1 Expression in the Bovine Corpus Luteum: Implications for Luteolysis Endocrinology, December 1, 2001; 142(12): 5254 - 5260. [Abstract] [Full Text] [PDF]
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C. FERNANDEZ-PATRON, C. ZOUKI, R. WHITTAL, J. S. D. CHAN, S. T. DAVIDGE, and J. G. FILEP Matrix metalloproteinases regulate neutrophil-endothelial cell adhesion through generation of endothelin-1[1-32] FASEB J, October 1, 2001; 15(12): 2230 - 2240. [Abstract] [Full Text] [PDF]
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 J. Yao, T. Morioka, B. Li, and T. Oite Endothelin is a potent inhibitor of matrix metalloproteinase-2 secretion and activation in rat mesangial cells Am J Physiol Renal Physiol, April 1, 2001; 280(4): F628 - F635. [Abstract] [Full Text] [PDF]
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 P. Jurasz, G. Sawicki, M. Duszyk, J. Sawicka, C. Miranda, I. Mayers, and M. W. Radomski Matrix Metalloproteinase 2 in Tumor Cell-induced Platelet Aggregation: Regulation by Nitric Oxide Cancer Res., January 1, 2001; 61(1): 376 - 382. [Abstract] [Full Text]
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S. Archer and S. Rich Primary Pulmonary Hypertension : A Vascular Biology and Translational Research "Work in Progress" Circulation, November 28, 2000; 102(22): 2781 - 2791. [Abstract] [Full Text] [PDF]
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 G. Sawicki, M. W. Radomski, B. Winkler-Lowen, A. Krzymien, and L. J. Guilbert Polarized Release of Matrix Metalloproteinase-2 and -9 from Cultured Human Placental Syncytiotrophoblasts1 Biol Reprod, November 1, 2000; 63(5): 1390 - 1395. [Abstract] [Full Text]
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C. Fernandez-Patron, K. G. Stewart, Y. Zhang, E. Koivunen, M. W. Radomski, and S. T. Davidge Vascular Matrix Metalloproteinase-2-Dependent Cleavage of Calcitonin Gene-Related Peptide Promotes Vasoconstriction Circ. Res., October 13, 2000; 87(8): 670 - 676. [Abstract] [Full Text] [PDF]
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 T. H. Vu and Z. Werb Matrix metalloproteinases: effectors of development and normal physiology Genes & Dev., September 1, 2000; 14(17): 2123 - 2133. [Full Text]
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 H. D. Intengan and E. L. Schiffrin Structure and Mechanical Properties of Resistance Arteries in Hypertension : Role of Adhesion Molecules and Extracellular Matrix Determinants Hypertension, September 1, 2000; 36(3): 312 - 318. [Abstract] [Full Text] [PDF]
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C. Fernandez-Patron, M. W. Radomski, and S. T. Davidge Role of matrix metalloproteinase-2 in thrombin-induced vasorelaxation of rat mesenteric arteries Am J Physiol Heart Circ Physiol, May 1, 2000; 278(5): H1473 - H1479. [Abstract] [Full Text] [PDF]
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 M. J. Sierevogel, E. Velema, P. P. de Jaegere, D. P. de Kleijn, C. Borst, and G. Pasterkamp Minimal Duration of Oral Matrix Metalloproteinase Inhibition to Prevent Constrictive Arterial Remodeling after Balloon Dilation in the Pig Radiology, February 1, 2002; 222(2): 468 - 473. [Abstract] [Full Text] [PDF]
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