|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
CHEMOKINES
From the Department of Biochemistry and Molecular
Biology, the Department of Oral Biological and Medical Sciences, and
the Biomedical Research Centre, University of British Columbia,
Vancouver, Canada; and the Department of Pharmacology and Therapeutics,
University of Calgary, Alberta, Canada.
Monocyte chemoattractant protein (MCP)-3 is
inactivated upon cleavage by the matrix metalloproteinase (MMP)
gelatinase A (MMP-2). We investigated the susceptibility to proteolytic
processing of the 4 human MCPs by 8 recombinant MMPs to determine
whether MCP-3 is an isolated example or represents a general
susceptibility of chemokines to proteolytic inactivation by these
important inflammatory proteases. In addition to MMP-2, MCP-3 is
efficiently cleaved by membrane type 1 (MT1)-MMP, the cellular
activator of MMP-2, and by collagenase-1 and collagenase-3 (MMP-1,
MMP-13) and stromelysin-1 (MMP-3). Specificity was shown by absence of
cleavage by matrilysin (MMP-7) and the leukocytic MMPs neutrophil
collagenase (MMP-8) and gelatinase B (MMP-9). The closely related
chemokines MCP-1, MCP-2, and MCP-4 were not cleaved by MMP-2 or
MT1-MMP, but were cleaved by MMP-1 and MMP-3 with varying efficiency.
MCPs were typically cleaved between residues 4 and 5, but MCP-4 was
further processed at Val7-Pro8. Synthetic MCP analogs
corresponding to the MMP-cleaved forms bound CC chemokine receptor
(CCR)-2 and CCR-3, but lacked chemoattractant activity in pre-B cells
transfected with CCR-2 and CCR-3 or in THP-1 monocytic cells, a
transformed leukemic cell line. Moreover, the truncated products of
MCP-2 and MCP-4, like MCP-3, were potent antagonists of their cognate CC chemokine receptors in transwell cell migration assays in vitro. When they were injected 24 hours after the initiation of
carrageenan-induced inflammation in rat paws, their in vivo
antagonist activities were revealed by a greater than 66% reduction in
inflammatory edema progression after 12 hours. We propose that
MMPs have an important role in modulating inflammatory and immune
responses by processing chemokines in wound healing and in disease.
(Blood. 2002;100:1160-1167) Chemokines are potent chemoattractant cytokines
that are produced locally in tissues and direct the migration and
homing of leukocytes. Tissue gradients of inflammatory chemokines
attract and maintain inflammatory cells at sites of host challenge in infection, inflammation, and cancer.1 Chemokines can be
divided into families according to the position and spacing of
N-terminal cysteine residues. Presently, the C, CC, CXC, and
CX3C families are recognized,2 with more than
54 human chemokines currently identified. The monocyte chemoattractant
proteins (MCPs) of the CC family consist of 4 proteins termed MCP-1,
MCP-2, MCP-3, and MCP-4 (CCL-2, CCL-8, CCL-7, and CCL-13, respectively)
that target multiple leukocyte subsets (monocytes, basophils,
eosinophils, dendritic cells, and natural killer cells) whereas, in the
initial phases of inflammation, CXC chemokines attract
polymorphonuclear leukocytes. Therefore, chemokines are important
mediators of many pathologies, including chronic inflammatory and
autoimmune diseases where the coordinated expression of MCPs and
resultant leukocyte infiltration correlate with disease
progression.3,4
An important question in the pathogenesis of inflammation and disease
is how chemoattractant signals are squelched to restrict new cell
recruitment and to promote clearance of the inflammatory infiltrate as
a prelude to tissue resolution. Proteolysis of chemokines may provide a
mechanism for loss of chemoattractant signaling. Indeed, production of
proteolytically processed chemokines in cell culture has been
reported.5-8 We have demonstrated an N-terminal-truncated form of MCP-3 in human rheumatoid synovial fluid that had been cleaved
at Gly4-Ile5, and the effects of engineered truncated chemokines on
cell behavior have been characterized.9 Hence, the
identification of proteinases that may process chemokines in vivo is
important, but as yet poorly characterized. Very recently, the matrix
metalloproteinase (MMP) gelatinase A (MMP-2) was shown to process the
N-terminus of MCP-3 and stromal cell-derived factor-1 MMPs are either secreted or cell-membrane-bound proteinases with broad
substrate specificity that have traditionally been proposed to degrade
most components of the extracellular matrix12 although in
vivo evidence for this is generally lacking. Matrix proteolysis is a
hallmark of inflammation with MMPs considered to be important effectors
of this process and also essential for leukocyte extravasation
and migration. In particular, leukocyte-derived collagenase-2 (MMP-8)
and gelatinase B (MMP-9) are the prominent early proteolytic mediators
of matrix degradation and allow effector cell egress to the site of
tissue damage.13,14 Following this initial leukocyte
proteolytic phase, stromal cells, in response to proinflammatory
cytokines secreted by the cellular infiltrate, produce MMPs that
amplify the acute tissue-destructive phase. Notably, interstitial
collagenase (MMP-1), stromelysin-1 (MMP-3), matrilysin (MMP-7), and
collagenase-3 (MMP-13) have been suggested to play important roles in
inflammatory tissue destruction.12 Hence, persistence of
the inflammatory infiltrate results in continued tissue destruction
that can progress to chronic inflammatory disease. Although MMPs may
play a role in extracellular matrix degradation in inflammation, it is
gradually becoming appreciated that these proteases have a wider
substrate repertoire that includes many bioactive molecules.
Indirectly, the MMP-mediated inactivation of serpins15,16
and chemokines7,8 can modulate healing with direct
biological effects of MMP cleavage manifested by the activation and
release of tumor necrosis factor- We have recently reported the use of the yeast 2-hybrid system to
screen for potential MMP substrates. Exosite scanning resulted in the
identification of MCP-3 as a binding protein and substrate for
MMP-2.7 This was a specific interaction, as MMP-2 did not cleave MCP-1, MCP-2, or MCP-4. We have also very recently identified SDF-1 Production of MMPs and MCPs
Electrospray ionization mass spectrometry
Proteinase assays P-aminophenylmercuric acetate (APMA)-activated MMP (1 ng) and 1 µg chemokine were mixed in buffer (100 mM NaCl, 5 mM CaCl2, 20 mM Tris, pH 8.0) and incubated at 37°C. Aliquots were removed at 1-hour intervals, and product accumulation was monitored by densitometric analysis of Coomassie-stained Tris-tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. The overall catalytic rate/Km (kcat/Km) specificity constant was calculated from graphical determination of the kobs.7 Electrospray ionization mass spectrometry of the reaction products was used to confirm cleavage, and the data were deconvoluted to identify the scissile bond.Chemotaxis assays Cell migration of THP-1 monocytic leukemia cells (ATCC, Manassas, VA) or a pre-B cell line (murine B300 19; ATCC) stably transfected with either human CCR-2 or human CCR-3 (designated B300-CCR2 and B300-CCR3, respectively) was evaluated in disposable 12-well Transwell polystyrene trays (Corning Costar, Cambridge, MA) across a polycarbonate membrane with 5-µm size pores. Full-length and N-terminal-truncated synthetic chemokines corresponding to the MMP-cleaved form of the chemokines were diluted in Hepes-buffered RPMI 1640 supplemented with 10 mg/mL bovine serum albumin. Chemokine samples were added to the lower chamber, and THP-1 cells or B19 300 cell transfectants (1 × 107 cells per milliliter), in the same media without the MCPs, were added to the upper chamber. After 3-hour incubation at 37°C in 5% CO2 humidified atmosphere, migrated cells in the lower chamber were counted. Migrated cells in 5 separate fields per well from duplicate wells were enumerated on a hemacytometer by means of light microscopy.125I-labeling of MCP-1 and measurement of chemokine receptor binding affinities The binding affinity (dissociation constant [Kd]) of MCP-2, MCP-2(5-76), MCP-4, and MCP-4(8-75) was determined by competition for 125I-labeled MCP-1 on CCR-2 and Scatchard analysis. The 125I-labeling of MCP-2 and MCP-4 resulted in precipitation of the chemokines, which could not be used for binding assays. MCP-1 (10 µg) was labeled with 9.25 × 105 Bq (250 µCi) 125I-Bolton Hunter reagent (NEN Life Science Products, Boston, MA) at 4°C for 30 minutes in 0.1 M borate buffer, and the reaction was terminated by incubation in 0.1 M glycine. The labeled MCP-1 was separated from free 125I-Bolton Hunter reagent by Sephadex G-25 size-exclusion chromatography. Binding assays were performed by means of CCR-2-transfected pre-B cells (5 × 106) suspended in 200 µL HEPES-buffered RPMI 1640 media, 10 mg/mL bovine serum albumin, and 0.1% NaN3. The 125I-MCP-1 (4 nM) and increasing concentrations of unlabeled competitor were added and incubated for 30 minutes. Cell-bound 125I-MCP-1 was separated from the unbound 125I-MCP-1 by pelleting the cells through a 2:3 mixture of diacetylphthalate and dibutylphthalate. The specifically bound counts per minute in the cell pellet were calculated after subtracting the nonspecifically bound counts per minute (counts per minute bound in the presence of 100-fold molar excess of unlabeled competitor) and dividing by the total counts per minute bound in the absence of competitor. Assays were performed in duplicate, and the experiments repeated.Rat paw inflammation model Male Wistar rats (200 to 225 g) (Charles River Breeding Farms, Montreal, QC, Canada) were housed in a biocontainment facility according to the Canadian Animal Care Committee with the use of institutionally approved animal care protocols. Inflammation was induced in rat paws by intraplantar injection of 1% lambda carrageenan (Sigma, St Louis, MO) in sterile saline (total volume, 100 µL) under halothane anesthesia. Paw volumes were measured by means of a hydroplethysmometer (Ugo Basile, Milan, Italy) before and 24 hours after injection of carrageenan and the induction of inflammation.27 After 24 hours, the rats were anesthetized with halothane, and an intraplantar injection of chemokine analogs in sterile saline (50 µg in 50 µL) or saline alone was administered according to a double-blind protocol. Five rats were used per treatment, and the experiment was repeated 2 or 3 times for each peptide. At 12 hours after peptide administration, paw volumes were measured in a randomized sequence by an observer unaware of the treatments. Changes in paw volume were then calculated by subtracting the final paw volume from the volume at the 24-hour time point.Statistical analysis Statistical analysis was performed by nonparametric (Kruskal-Wallis) analysis of variance followed by a Dunn multiple comparison test.
MCP-3 is cleaved by multiple MMPs The specificity of 8 MMPs that have been associated with wound healing12,28 in the proteolysis of MCP-3 was determined. Incubation of the chemokine with recombinant MMP-1, MMP-2, MMP-3, MMP-13, and MT1-MMP resulted in a small but distinct increase in electrophoretic mobility on Tris-tricine gels, whereas MMP-7, MMP-8, and MMP-9 did not, even with prolonged incubation (Figure 1). Cleavage of full-length chemokine was confirmed by electrospray ionization mass spectroscopic identification of the new lower molecular mass product (Figure 1). Deconvolution of the mass spectrometry data revealed that the MCP-3 scissile bond for each proteolytically competent MMP was Gly4-Ile5 (Table 1). We designated this cleavage product MCP-3(5-76). MMP-mediated processing of MCP-3 was efficient, as shown by the high turnover rates (Table 1), with MMP-2 and MT1-MMP showing highest activity.
Multiple MMPs cleave MCP-1, MCP-2, and MCP-4 We tested the ability of the MMPs to cleave MCP-1, MCP-2, and MCP-4. As found before,7 MMP-2 was highly specific for MCP-3 with no activity against MCP-1, MCP-2, or MCP-4. Figure 2 shows that MCP-1 is cleaved at Ala4-Ile5, designated MCP-1(5-76); by MMP-1; to a lesser extent (20% of total by densitometric analysis) by MMP-3; and to a very minor extent (5% of total) by MMP-8. In contrast, MCP-2 was susceptible to cleavage only by MMP-3 at Ser4-Val5, designated MCP-2(5-76). Interestingly, MCP-4 was processed at 2 sites, Asp3-Ala4 and Ala4-Leu5, by MMP-3. Both sites were also susceptible to MMP-1 activity, with proteolysis proceeding by a further cleavage at Val7-Pro8, designated MCP-4(8-75). Cleavage site susceptibility for MCP-1, MCP-2, MCP-3, and MCP-4 is summarized schematically in Figure 3.
MCP-2(5-76) is a CCR-2 receptor antagonist To compare the biological activity of full-length MCP-2 and MCP-2(5-76), we performed chemotaxis assays using synthetically prepared MCP-2(5-76) and a pre-B cell line stably transfected with the predominant MCP-2 chemokine receptor CCR-2. Figure 4A shows that, unlike full-length MCP-2, MCP-2(5-76) was inactive as a CCR-2 agonist at all concentrations up to 10 6 M in the chemotaxis assay. However, MCP-2(5-76)
retained receptor-binding affinity (Kd, 4.5 nM)
very similar to that for MCP-2 (Kd, 4.4 nM)
(Figure 5) and was effective as a CCR-2
antagonist in cell migration assays (Figure 4B).
MCP-4(8-75) is a CCR-2 and CCR-3 receptor antagonist The agonist and antagonist activity of MCP-4(8-75) was compared with that of MCP-4 with the use of 2 pre-B cell lines stably transfected with either CCR-2 or CCR-3, to which full-length MCP-4 binds with high affinity. Compared with the full-length chemokine, MCP-4(8-75) was unable to direct chemotaxis in either cell line (Figure 6A,B). Like the MMP-cleaved forms of MCP-2 and MCP-3, MCP-4(8-75) was a potent receptor antagonist of both CCR-2 and CCR-3 (Figure 6C,D). MCP-4(8-75) was calculated to bind CCR-2 with a Kd of 14 nM, an affinity very similar to that of the full-length form (Kd, 16 nM) (Figure 5).
THP-1 cell migration in response to MCP-2(5-76) and MCP-4(8-75) As a model of inflammatory monocytes, we used THP-1 monocytic cells, a transformed leukemic cell line that expresses CCR-2, to further assess the effects of MMP cleavage of MCP-2 and MCP-4. Whereas the full-length MCPs directed chemotaxis across transwell filters, MCP-2(5-76) and MCP-4(8-75) showed no chemotactic activity (Figure 7A,B), but both were receptor antagonists as evident by the dose-dependent reduction in THP-1 cell chemotaxis (Figure 7C,D), confirming the previous results using the pre-B cells transfected with CCR-2.
MMP-cleaved MCPs reduce inflammatory edema in vivo To determine the biological effects of MMP cleavage of MCPs in vivo, the extent of inflammatory edema induced in a rat paw model following injection with carrageenan was measured after administration of synthetic N-terminal-truncated MCPs corresponding to the MMP-cleaved forms. Results are shown in Figure 8 as the change in paw volume from the 24-hour time point, when the chemokines were injected, to the 36-hour postcarrageenan time point. Without chemokine, the paw volume continued to increase from the 24-hour value (1.80 ± 0.02 mL) as the inflammatory response progressed. MCP-1(5-76) and MCP-4(8-75) showed an approximately 66% reduction in the increase in inflammatory edema from 24 to 36 hours compared with the vehicle control (Figure 8). An engineered form of MCP-1(9-76), a potent CCR-2 antagonist,9 was found to reduce the paw volume to a similar extent. Most notable though was MCP-3(5-76), which reduced the paw volume 1.6-fold to below that at the time of injection. Hence, MMP inactivation of MCPs also generates CCR antagonists that retain cellular binding affinity and that modulate cell migration in vitro and dampen the inflammatory response to carrageenan in vivo.
MMPs are implicated in many physiological processes involving
matrix turnover,12,28 but direct evidence for this is
limited.29 In contrast, MMPs are clearly identified with
pathologies, including arthritis and tumor metastasis30
where they have been assumed to primarily play a matrix degradative
role. However, a broader MMP substrate degradome is now being revealed
by recent studies (reviewed in Overall31) that show
proteolytic susceptibility of signaling proteins such as
MCP-3,7 SDF-1 Because MMPs are differentially expressed in many tissues, in many cell
types, and during inflammation and healing, we determined the
specificity of 8 MMPs against the CC chemokines MCP-1, MCP-2, MCP-3,
and MCP-4. In addition to MMP-2, MMP-1, MMP-3, MMP-8, MMP-13, and
MT1-MMP cleave MCPs and all characteristically perform this at position
4-5, but the MMPs are not always functionally interchangeable and each
MCP showed a different profile of proteolytic susceptibility. The
pattern of MMP proteolysis of the MCP family is striking. MMP-3, a
protease with broad extracellular matrix substrate specificity (see
www.clip.ubc.ca/mmps.shtm for a comprehensive updated list of MMP
substrates), can cleave all MCP chemokines, but MMP-2, MMP-13, and
MT1-MMP are active only on MCP-3. The similar specificity of MMP-2 and
MT1-MMP is interesting because MT1-MMP is the major cellular activator
of proMMP-2, and both proteases assemble and function together in
activation complexes on the cell surface.25,33,35 Most
notable was the general inability of MMP-7 and of the primarily leukocytic enzymes MMP-8 and MMP-9 to process MCPs. Although MMP-8 could cleave MCP-1, catalytic efficiency was very low. MMP-9 can activate IL-8 by N-terminal processing, but does not cleave
MCP-2,36 or any other MCP as shown here. It is important
to distinguish between efficient and precise proteolytic processing, as
shown for these chemokines, and the more general catabolic actions of proteases during protein degradation. MMP-9 has been reported to slowly
degrade the chemokines PF-4, GRO- In addition to the amino acid sequence of the substrate, auxiliary
elements of substrate specificity are pivotally important, including
substrate binding by exosites on the MMP hemopexin C domain.7,31 The absence of a hemopexin C domain in MMP-7
may explain the lack of activity against these chemokines, and the hemopexin C domain of MMP-9 is suggested to be positioned away from the
catalytic domain by an extended carbohydrate-rich linker peptide,34 possibly accounting for its lack of ability to
process the N-terminus of the MCPs. Therefore, the same position of the MMP cleavage site in MCPs reported here and SDF-1 Neoepitope antibodies have been used to show the generation of
MCP-3(5-76) in human arthritis in vivo.7 Processing of
SDF-1 Previous protein engineering studies9,26,32 have demonstrated that modification of the N-terminus of MCP chemokines alters receptor binding and activation. In this report, we have shown that MMP processing of MCP-1, MCP-2, MCP-3, and MCP-4 reduces cell migration of THP-1 monocytic leukemia cells and pre-B cells transfected with CCR-1, CCR-2, and CCR-3. Cell surface receptor binding of the cleaved chemokines still occurred with only slightly altered Kds. Thus, MMP-processing of MCP-2, MCP-3, and MCP-4 generated stable and potent CCR-1, CCR-2, and CCR-3 receptor antagonists. The antagonistic effects are due to competition for receptor occupancy, with subsequent desensitization the result of ligated receptor trafficking from the cell surface to endosomes.9 Although primary cultures of monocytes were not tested for the effects of MMP cleavage on chemotaxis, the potential for the in vivo generation of N-terminally truncated forms of MCPs by MMPs that may modify inflammatory and immune responses was demonstrated with the use of THP-1 monocytic cells. Our previous characterization of synthetic MCP-1(5-76),9 corresponding to the MMP-cleaved form generated here, revealed that MCP-1(5-76) exhibited a 10-fold reduction in receptor agonist activity. MCP-1(5-76) bound CCR-2 with a Kd of 20 nM and desensitized the receptor to subsequent chemokine treatment.9 Although MCP-1(5-76) can be detected as a weak agonist in sensitive in vitro assays, this can be outweighed in vivo by its antagonistic properties, as evident by the reduction in inflammatory edema 12 hours after its injection into inflamed paw pads. Comparable effects were found for MCP-4(8-75) and for MCP-2(5-76). The effectiveness of MCP-3(5-76), a broad-spectrum CCR antagonist, in reducing inflammation in vivo in other models7 previously suggested the potential importance of generating a broad-spectrum chemotactic antagonist for CCR-1, CCR-2, and CCR-3 in modulating inflammatory processes. Our present data support this hypothesis. Compared with the other MCP antagonists, which cover a more restricted CCR spectrum, MCP-3(5-76) was found to have the strongest anti-inflammatory effects, reducing the paw volume to below that at the time of injection: MCP-3(5-76) not only prevented new edema, but resolved some of the pre-existing inflammatory exudate present after 24 hours of inflammation. The cleavage and conversion of MCP-1, MCP-2, and MCP-4 to additional antagonists are likely to augment this response in different diseases and processes. Hence, the pathophysiological cleavage of MCPs by MMPs reduces CCR agonist activity and generates effective antagonist derivatives that may regulate inflammatory and immune processes in vivo. However, whether a connection exists between MMP activity and the modulation of CCR-1-, CCR-2-, and CCR-3-dependent cellular responses in disease remains to be elucidated and is currently under investigation in our laboratories. The physiological likelihood of in vivo chemokine cleavage, specifically MCP-3, was revealed by the high kinetic turnover rates (Table 1). Notably, MMP-2 and MMP-14 were the most efficient at cleaving MCP-3 in vitro, but interestingly neither cleaved MCP-1, MCP-2, or MCP-4. This points to a unique role for MCP-3 cleavage by MT1-MMP and MMP-2. MT1-MMP is the physiological activator of MMP-2 and forms a cell-surface receptor for this enzyme.35 MT-MMPs are also critical for cell migration in collagen,37 an important feature of remodeling wounds. We have previously postulated that modulation of the MMP-2/MT1-MMP proteolytic axis at the cell surface changes the proteolytic profile of stromal cells from collagenolytic (predominantly manifested by MT1-MMP activity) to gelatinolytic (by MMP-2) during the conversion from a cytokine-stimulated resorptive cell to a matrix-depositing cell.31,33 Proteolysis of MCP-3 by MT1-MMP and MMP-2 recruited to the cell surface in both the collagenolytic and gelatinolytic phases of tissue remodeling is likely to also be augmented by the action of collagenase and stromelysin, which may process MCP-1, MCP-2, and MCP-4 during the tissue resorptive phase, to reduce chemokine activity directly and indirectly by creating CCR-1, CCR-2, and CCR-3 antagonist gradients. Although TIMPs present in the tissue reduce net proteolytic activity by MMPs, particularly at the interface between the inflammatory lesion and normal tissue, TIMP-2 binding to the hemopexin C domain of MMP-2 interferes with neither MCP-3 binding and cleavage nor MMP-2 activation by MT1-MMP (G.A.M., C.M.O., unpublished data, May 2000). Our data support the following model (Figure
9), which connects the activity of
chemokines and MMPs in the stages that define the inflammatory
reaction. Chemoattractant-directed leukocytes secrete MMPs,
predominantly MMP-8 and MMP-9 but not MMP-2,38-40 that may
degrade matrix and promote migration but do not cleave MCPs. As the
inflammatory reaction progresses, cytokines such as interleukin-1 and
TNF-
The present and recent data from our group7 showing the
efficient cleavage of MCPs 1, 2, 3, and 4 and SDF-1
We thank Shouming He for technical assistance in mass spectrometry analysis.
Submitted July 3, 2001; accepted April 12, 2002.
Supported by grants from the Canadian Arthritis Network of Centers of Excellence, the National Cancer Institute of Canada with funds provided in part by the Canadian Cancer Society, and the Canadian Institutes for Health Research. G.A.M. is supported by a National Cancer Institute of Canada Studentship; I.C.-L. is supported by a Canadian Institutes for Health Research Scientist Award; and C.M.O. is supported by a Canadian Research Chair in Metalloproteinase Biology.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Christopher M. Overall, 2199 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada; e-mail: chris.overall{at}ubc.ca.
1.
Foxman EF, Campbell JJ, Butcher EC.
Multistep navigation and the combinatorial control of leukocyte chemotaxis.
J Cell Biol.
1997;139:1349-1360 2. Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity. Immunity. 2000;12:121-127[CrossRef][Medline] [Order article via Infotrieve]. 3. Schluger NW, Rom WN. Early responses to infection: chemokines as mediators of inflammation. Curr Opin Immunol. 1997;9:504-508[CrossRef][Medline] [Order article via Infotrieve]. 4. Baggiolini M. Chemokines and leukocyte traffic. Nature. 1998;392:565-568[CrossRef][Medline] [Order article via Infotrieve]. 5. Proost P, Struyf S, Wuyts A, et al. Isolation and identification of naturally modified C-C chemokines MCP-1, MCP-2 and RANTES: effects of posttranslation modifications on receptor usage, chemotactic and anti-HIV-1 activity. Eur Cytokine Netw. 1998;9:73-75[Medline] [Order article via Infotrieve]. 6. Wuyts A, Govaerts C, Struyf S, et al. Isolation of the CXC chemokines ENA-78, GRO alpha and GRO gamma from tumor cells and leukocytes reveals NH2-terminal heterogeneity: functional comparison of different natural isoforms. Eur J Biochem. 1999;260:421-429[Medline] [Order article via Infotrieve].
7.
McQuibban GA, Gong JH, Tam EM, McCulloch CA, Clark-Lewis I, Overall CM.
Inflammation dampened by gelatinase A cleavage of monocyte chemoattractant protein-3.
Science.
2000;289:1202-1206
8.
McQuibban GA, Butler GS, Gong JH, et al.
Matrix metalloproteinase activity inactivates the CXC chemokine stromal cell-derived factor-1.
J Biol Chem.
2001;276:43503-43508
9.
Gong JH, Clark-Lewis I.
Antagonists of monocyte chemoattractant protein 1 identified by modification of functionally critical NH2-terminal residues.
J Exp Med.
1995;181:631-640 10. Van Damme J, Struyf S, Wuyts A, et al. The role of CD26/DPP IV in chemokine processing. Chem Immunol. 1999;72:42-56[Medline] [Order article via Infotrieve]. 11. Delgado MB, Clark-Lewis I, Loetscher P, et al. Rapid inactivation of stromal cell-derived factor-1 by cathpsin G associated with lymphocytes. Eur J Immunol. 2001;31:699-707[CrossRef][Medline] [Order article via Infotrieve].
12.
Nagase H, Woessner JF.
Matrix metalloproteinases.
J Biol Chem.
1999;274:21491-21494 13. D'Haese A, Wuyts A, Dillen C, et al. In vivo neutrophil recruitment by granulocyte chemotactic protein-2 is assisted by gelatinase B/MMP-9 in the mouse. J Interferon Cytokine Res. 2000;20:667-674[CrossRef][Medline] [Order article via Infotrieve]. 14. Madri JA, Graesser D. Cell migration in the immune system: the evolving inter-related roles of adhesion molecules and proteinases. Dev Immunol. 2000;7:102-116.
15.
Mast AE, Enghild JJ, Nagase H, Suzuki K, Pizzo SV, Salvesen G.
Kinetics and physiologic relevance of the inactivation of alpha 1-proteinase inhibitor, alpha 1-antichymotrypsin, and antithrombin III by matrix metalloproteinases-1 (tissue collagenase), -2 (72-kDa gelatinase/type I collagenase), and -3 (stromelysin).
J Biol Chem.
1991;266:15810-15816 16. Liu Z, Zhou X, Shapiro SD, et al. The serpin alpha1-proteinase inhibitor is a critical substrate for gelatinase B/MMP-9 in vivo. Cell. 2000;102:647-655[CrossRef][Medline] [Order article via Infotrieve]. 17. Gearing AJ, Beckett P, Christodoulou M, et al. Processing of tumor necrosis factor-alpha precursor by metalloproteinases. Nature. 1994;370:555-557[CrossRef][Medline] [Order article via Infotrieve].
18.
Levi E, Fridman R, Miao HQ, Ma YS, Yayon A, Vlodavsky I.
Matrix metalloproteinase 2 releases active soluble ectodomain of fibroblast growth factor receptor 1.
Proc Natl Acad Sci U S A.
1996;93:7069-7074 19. Powell WC, Fingleton B, Wilson CL, Boothby M, Matrisian LM. The metalloproteinase matrilysin (MMP-7) proteolytically generates active soluble Fas ligand and potentiates epithelial cell apoptosis. Curr Biol. 1999;9:1441-1447[CrossRef][Medline] [Order article via Infotrieve]. 20. Fingleton B, Vargo T, Crawford HC, Matrisian LM. Matrilysin expression selects for cells with reduced sensitivity to apoptosis. Neoplasia. 2001;3:459-468[CrossRef][Medline] [Order article via Infotrieve].
21.
Wilson CL, Ouellette AJ, Satchell DP, et al.
Regulation of intestinal alpha-defensin activation by the metalloproteinase matrilysin in innate host defense.
Science.
1999;286:113-117
22.
Butler GS, Sim D, Tam E, Devine E, Overall CM.
Mannose binding lectin (MBL) mutants are susceptible to matrix metalloproteinase proteolysis: potential role in MBL deficiency.
J Biol Chem.
2002;277:17511-17519
23.
Van den Steen PE, Proost P, Wuyts A, Van Damme J, Opdenakker G.
Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoterminal processing, whereas it degrades CTAP-III, PF-4, and GRO- 24. Barret AJ. Proteolytic Enzymes: Aspartic and Metallo Peptidases. San Diego, CA: Academic; 1995.
25.
Bigg HF, Morrison CJ, Butler GS, et al.
Tissue inhibitor of metalloproteinases-4 (TIMP-4) inhibits, but does not support, the activation of gelatinase A via efficient inhibition of membrane type 1-matrix metalloproteinase.
Cancer Res.
2001;61:3610-3618
26.
Gong JH, Uguccioni M, Dewald B, Baggiolini M, Clark-Lewis I.
RANTES and MCP-3 antagonists bind multiple chemokine receptors.
J Biol Chem.
1996;271:10521-10527 |
Top
Abstract
Introduction
(SDF-1) and
.
ie, with 0 M MCP-4(8-75). (D) Cell migration
assay of B300-CCR3 in response, as indicated, to full-length MCP-4 (100 nM) and in the presence of increasing concentrations of MCP-4(8-75) or
MCP-4 alone, ie, with 0 M MCP-4(8-75). Mean values of
duplicate assays are shown.
