From the Institute of Enzymology, Biological Research
Center, Hungarian Academy of Sciences, Budapest H-1518, Hungary and
§ Department of Chemistry, Carnegie Mellon University,
Pittsburgh, Pennsylvania 15213
Received for publication, October 2, 2002, and in revised form, November 20, 2002
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ABSTRACT |
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The interaction of matrix
metalloproteinase 2 (MMP-2) with gelatin is mediated by three repeats
homologous to fibronectin type II (FN2) modules, which are inserted in
the catalytic domain in proximity of the active site. We screened a
random 15-mer phage display library to identify peptides that interact
with the FN2 modules of MMP-2. Interestingly, the selected peptides are
not gelatin-like and do not share a common, obvious sequence motif. However, they contain a high proportion of aromatic residues. The
interactions of two peptides, WHWRH0RIPLQLAAGR and
THSHQWRHHQFPAPT, with constructs comprising the in-tandem first
and second and second and third FN2 modules of MMP-2 (Col-12 and
Col-23, respectively) were characterized by NMR. Both peptides interact
with Col-12 and Col-23 with apparent association constants in the
mM Matrix metalloproteinase 2 (MMP-2,1 gelatinase A), and
the closely related MMP-9 (gelatinase B)
are unique among the metalloproteinases in that three gelatin-binding
fibronectin type II (FN2) modules (Col-1, Col-2, and Col-3) are
inserted in their catalytic domain in the vicinity of the active site
(1). The solution conformation of each FN2 repeat from human MMP-2 has
been characterized via NMR spectroscopy (2-4). Moreover, the x-ray
crystallographic structure of the intact human pro-MMP-2 has been
reported (5).
In the second FN2 module from each MMP-2 and MMP-9, residues that are
important for the interaction with gelatin have been identified via
site-directed mutagenesis (6, 7). Additionally, the ligand binding
surfaces of all three modules of MMP-2 have been mapped from
1H and 15N NMR perturbations induced by
(PPG)6 and the longer chain analog, (PPG)12,
synthetic peptide mimics of gelatin (2-4). In line with the
crystallographic evidence, which shows that the FN2 modules in MMP-2
point away from each other (5), our NMR studies of the interaction
between Col domains and (PPG)6 and (PPG)12 have shown that consecutive Col modules contain distinct ligand-binding sites in which affinities for these ligands are virtually identical to
those of the individual domains (3, 4, 8).
Although the affinity of the MMP-2 Col domains for collagenous ligands
appears by now to be well established, less is known regarding the
specificity of the interaction. In our previous studies we found that
the peptide PIIKFPGDVA, which corresponds to segment 33-42 of the
pro-MMP-2, interacts with the three Col domains of MMP-2 in a manner
that mimics the interaction with the collagen-like (PPG)6
and (PPG)12 peptides (3, 4). Preference for binding to
Col-3 was indicated, consistent with the x-ray crystallographic
structure of the pro-MMP-2 (5). In the proenzyme, the prodomain
interacts intramolecularly with the putative gelatin-binding site of
Col-3 via contacts that involve propeptide amino acid residues
Ile-35, Phe-37, and Asp-40. As these studies indicate, the
ligand specificity of the Col domains is not restricted to collagen-like peptides. It would be useful to gain more information as
to the range of structural diversity acceptable for peptides to
interact with the Col-binding sites.
In the context of MMP-2 involvement in tumor invasion, metastasis and
other physiopathological processes (reviewed in Ref. 9), it is highly
desirable to identify agents that could block its activity. The
suitability of MMP-2 as an anticancer target is supported by the
finding that MMP-2-deficient mice display reduced angiogenesis and
tumor progression (10). However, very few inhibitors specific for MMP-2
have been described to date (11). The most potent inhibitors also
inhibit several other MMP family members (12-14). Although generic MMP
inhibitors prevent tumor dissemination and formation of metastases in
animal models (14-19), they tend to elicit too broad a spectrum
of response and often exhibit side effects. It can be speculated that
active site inhibitors that also bind to the unique FN2 domains of
MMP-2 may be more MMP-2-specific. As a platform for such studies, we
have screened random 6-mer and random 15-mer phage display libraries for peptides that interact with the FN2 domains of MMP-2 and
characterized the interaction of two selected peptides with the
homologous gelatin-binding repeats via 1H/15N
NMR studies on the Col-12 and Col-23 constructs.
Selection of Peptides from Phage Display
Libraries--
Microtiter plates (Greiner Labortechnik) were coated
with the first (
Two phage fUSE5 libraries, which express a foreign 15- or 6-mer random
peptide library at the amino-terminal end of all five copies of its
pIII coat protein (22) and the Escherichia coli strain
K91Kan (thi/HfrC), carrying a "mini-kan hopper" element inserted in
the lacZ gene, were obtained from Prof. G. Smith
(University of Missouri-Columbia). The number of primary clones in the
15-mer library is 2.5 × 108 (23), and in the case of
the 6-mer library it is 2 × 108 (22). Approximately
1010 phage/well in 100 µl of TBS buffer were
incubated for 60 min at room temperature. Nonspecifically adsorbed
phage were washed away with 12 times with 200 µl of TBS buffer
containing 5 mg/ml serum albumin and 0.5% Tween 20. Unless otherwise
indicated, bound phage were eluted with 2 times with 200 µl of
TBS buffer containing 1 mg/ml gelatin type A from porcine skin type I
collagen (Sigma). In some experiments with immobilized
The peptides ACGYTYHPPCARLTV (ACG), WFPGPITFIPRPWSS (WFP),
WHWRHRIPLQLAAGR (WHW), THSHQWRHHQFPAPT (THS), and HASHFRFRHSHVYGV (HAS)
were synthesized on an AB-PE 431A peptide synthesizer (Applied Biosystems-PerkinElmer Life Sciences) using Fmoc
(N-(9-fluorenyl)methoxycarbonyl) chemistry.
NMR-monitored Peptide Binding--
15N-labeled
Col-12 and Col-23 modules from human MMP-2 (residues 223-337 and
278-394 respectively) (Fig. 1) were expressed in E. coli
and purified as described previously (3, 4).
To monitor ligand-induced resonance shifts, small aliquots
of WHW or THS stock solutions in 90% H2O, 10%
D2O, pH 7.0, were added to samples of 0.35 mM
15N-labeled Col-12 and Col-23 in 90% H2O, 10%
D2O, pH 7.0, and 1H-15N HSQC
experiments (25-27) were recorded at each step. All of the data were
acquired at 25 °C on Bruker Avance DMX-500 spectrometer equipped
with a 5-mm triple resonance three-axis gradient probe. The spectra
were processed and analyzed with the programs Felix 95 and Felix 98 (Molecular Simulations, Inc., San Diego, CA) on a Silicon Graphics Indy
R-5000 work station. Protein and peptide ligand concentrations were
determined spectrophotometrically (28). Values of the equilibrium
association constant (Ka) were determined by a
combination of linear and nonlinear least squares fitting of the
chemical shift changes, as described previously (29, 30).
Selection of Peptides That Interact with FN2 Domains from
MMP-2--
To identify peptides that bind to the FN2 modules from
MMP-2, phage display 6-mer or 15-mer random peptide libraries were screened with
In the case of the 6-mer library-unlike in the case of the 15-mer
library-there was no enrichment of bound phage after three rounds of
biopanning. This observation suggested that a six-residue-long peptide
may be too short for unique recognition of FN2 domains, therefore we
concentrated on the 15-mer library. From the latter library, after
three rounds of biopanning, individual clones were sequenced and the
following peptides were identified: ACGYTYHPPCARLTV (ACG),
WFPGPITFIPRPWSS (WFP), WHWRHRIPLQLAAGR (WHW), THSHQWRHHQFPAPT (THS), WHVSPRHQRLFHGLF (WHV) and HASHFRFRHSHVYGV (HAS). The
frequencies of the peptides selected under various conditions are
summarized in Table I. Interestingly, the
peptides are not collagen-like and do not share a common, obvious
sequence motif. However, their sequences exhibit biased amino
acid composition. For example, although His accounts for only 2% of
residues in protein databases, the selected peptides contain 16% His.
This trend is even more pronounced for peptides selected on single FN2
domains, which contain 22% His. There also is bias in the aromatic
amino acid content: whereas Tyr, Trp and Phe account for 8% of
residues in protein databases, in the selected peptides, their
proportion is 16%. Interestingly, peptide inhibitors of the whole
gelatinases that have been selected previously from phage display
library (11) are similarly enriched in aromatic residues whereas those from combinatorial (31) library contain multiple histidines. Our
results suggest that not only interaction with the active site but also
binding to FN2 modules may contribute to activity of these inhibitors.
It is also noteworthy that peptide WFP contains sequence FPG which is
found in the pro-domain (residues 37-39); F37 from this motif inserts
into the hydrophobic gelatin-binding pocket of Col-3 in the x-ray
structure of pro-MMP-2 (5).
The clones selected on
The ACG peptide, which was recovered with the greatest frequency on
immobilized
There is a striking difference between clones eluted from NMR Characterization of the Interaction between FN2 Modules and
Selected Peptides--
The interaction of the peptides with Col-12 and
Col-23 was investigated using NMR (Fig.
1). Because of solubility problems, only
WHW and THS were found suitable for this study. Col-12 and Col-23
chemical shift changes induced by WHW and THS binding were monitored in
1H-15N HSQC spectra (Fig.
2), and affinity constants
(Ka) were calculated from the ligand titration
curves (Fig. 3, Table II). The first and
second modules of the Col-12 construct bind to WHW with
Ka ~ 1.9 ± 0.2 mM
Previously (2-4), we mapped the gelatin binding surface of the Col
modules by localizing spectral perturbations induced by the synthetic
gelatin-like peptides (PPG)6 and (PPG)12 on the three-dimensional structures of the modules (Fig.
4). This approach proved to be less
straightforward in the current study. Backbone amide resonances
stemming from the termini and the linking segment of the two-domain
constructs are affected by WHW (Figs. 2
and 5) and THS (Fig. 6); similar effects
were observed during titration of Col-23 with the synthetic gelatin
mimics (compare against Fig. 4, C and D).
However, in the latter study, the shifts arising from the linking
peptide were negligible relative to those marking the gelatin-binding
pocket. In the current experiments, on the other hand, most spectral
perturbations were of similar magnitude or smaller than those localized
in the termini and linker. Hence, it is difficult to distinguish the
effects caused by direct contact with the ligand from shifts
related to altered conformation or dynamics due to peptide binding
elsewhere. Chemical shift perturbations at locations distant from
ligand contact sites have been observed previously in other systems
(35).
The gelatin-binding surfaces of Col modules comprise an aromatic
cluster and its surrounding region, in particular the loop comprising residues 33-38, at the front face of the domains. The distribution of WHW- and THS-induced shifts on the three-dimensional structures of Col modules can be summarized as follows (Figs. 5 and 6).
In the first FN2 module within the Col-12 construct (Col-12/1), WHW
predominantly affects residues neighboring the gelatin-binding pocket,
most notably residues Gly-33, Arg-34 and Trp-40. In the second FN2
module within both the Col-12 and Col-23 constructs (Col-12/2 and
Col-23/2), WHW perturbs mainly the termini, whereas the loop on the
front face, which is involved in gelatin binding, is affected to a
lesser extent. In the third FN2 module in Col-23 (Col-23/3),
WHW-induced resonance shifts are limited to the back of the domain and
include the termini and exposed hydrophobic patch, which encompasses,
among others, residues Cys-15 to Phe-17; the front side is affected
only minimally. Thus, it may be concluded that WHW interacts with the
gelatin-binding pocket of Col-1 (4) and possibly Col-2. In agreement
with such an interpretation, WHW has been selected most frequently by
gelatin elution from plates coated with Design of inhibitors that act on MMP-2 but not on other
metalloproteases is a challenging task. Because MMP-2 (together with MMP-9) is unique in having FN2 domains positioned next to its catalytic
cleft, active site inhibitors that also interact with FN2 domains
should be much more specific. Hence, we have screened random 6- and
15-mer phage display libraries and identified several peptides from the
latter library that interact with the FN2 modules of MMP-2.
The selected peptides were found to contain a high proportion of
aromatic residues and no acidic side chains. Enrichment in aromatic
amino acids may reflect the fact that such peptides are more likely to
have a fixed conformation and thus may have higher affinity than more
flexible peptides. The same reasoning may explain why no significant
enrichment was observed in the case of shorter peptides.
The interaction of two peptides, WHW and THS, with FN2 modules was
characterized by NMR. Both peptides bound to the Col domains with
Ka in the mM It is a well established strategy in drug design to chemically link two
low-affinity ligands to generate a high-affinity, high-specificity
compound (36). As an application, peptides binding to FN2 modules can
be linked to active site inhibitors with the aim of significantly
increasing the affinity and specificity of the interaction. It is our
hope that the peptide ligands we have identified will provide useful
leads for the development of more potent gelatinase inhibitors.
1 range. Peptide binding results in
perturbation of signals from residues located in the gelatin-binding
pocket and flexible parts of the molecule. Although the former
finding suggests that the gelatin-binding site is involved in the
contact, the interpretation of the latter is less straightforward and
may well reflect both the direct and indirect effects of the interaction.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
CONCLUSIONS
REFERENCES
MATERIALS AND METHODS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
CONCLUSIONS
REFERENCES
galCol-1), second (
galCol-2), or third
(
galCol-3) FN2 modules from human MMP-2 (20) or with the three
domains in tandem (
galCol-123) (21). The recombinant proteins,
consisting of the appropriate FN2 module(s) and an amino-terminal
peptide derived from the
-galactosidase moiety of the expression
vector, were prepared as described previously (20, 21). The plates were
incubated with the proteins (20 µg/ml) in 100 mM
NaHCO3 buffer for 2 h at 37 °C, after which they
were blocked with 30 mg/ml serum albumin in 100 mM
NaHCO3 buffer for 2 h at 37 °C and washed six times
with TBS buffer (50 mM Tris-HCl, pH 7.5, containing 150 mM NaCl) and 0.5% Tween 20.
galCol-123-Sepharose was prepared using cyanogen bromide-activated
Sepharose 4B (Amersham Biosciences) and
galCol-123 according to
instructions from the manufacturer.
galCol-123, elution was performed with buffer containing 2 mg/ml
galCol-123. Alternatively, ~1010 phage in 200 µl of
TBS buffer were incubated with 50 µl of
galCol-123-Sepharose for
60 min. The resin was then washed with 10 ml of TBS buffer containing 5 mg/ml serum albumin and 0.5% Tween 20, and the bound phage were eluted
with 3 × 200 µl of TBS buffer containing 1 mg/ml gelatin. The
eluted phages were amplified, and a portion was used in the next
biopanning cycle (24). After three rounds, individual phage were
isolated, and the DNA sequence of the 5' end of the gene III was
determined using a primer complementary to the positions 1663-1680 of
the wild type gene.
RESULTS AND DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
CONCLUSIONS
REFERENCES
galCol-1,
galCol-2,
galCol-3 and
galCol-123. Nonspecifically adsorbed phage were washed away with buffer containing 5 mg/ml serum albumin, then-in most experiments-specifically bound phage was eluted with buffer containing gelatin.
Frequency (%) of phage display peptides selected under various
conditions
galCol-1,
galCol-2 and
galCol-3 exhibit
identical sequences with only moderate differences in their relative
distributions. In turn, this indicates that despite the sequence
differences between the modules, the ligand-binding sites of the three
domains are likely to possess common features.
galCol-123, has two cysteines that may form a cystine
bridge, thus constraining the structure. It is a common observation
with phage display peptides that those with the highest affinity tend
to be cyclic (11, 32-34). Interestingly, ACG was rarely selected on
single Col domains, which suggests that the complementary binding
surface on
galCol-123 comprises multiple modules.
galCol-123
with gelatin and those eluted with
galCol-123. Although gelatin will
primarily elute phage that interacts with the gelatin-binding site of Col-123,
galCol-123 is likely to release phage bound to any
part of the protein. Therefore, peptides that become enriched upon
elution with
galCol-123 relative to elution with gelatin (WHV and,
to a lesser extent, WHW) are likely to interact with a region situated
at least partially outside of the gelatin-binding surface.
1
(Col-12/1) and Ka ~ 2.6 ± 0.3 mM
1 (Col-12/2).
Within experimental errors, the latter agrees with the value derived
for Col-23/2, Ka ~ 2.8 ± 0.4 mM
1. For Col-23/3, Ka ~ 3.0 ± 0.5 mM
1 was determined. The
affinity of THS for the Col modules is somewhat weaker, with
Ka ~ 0.9 ± 0.2 mM
1
for Col-12/1, Ka ~ 0.9 ± 0.3 mM
1 for Col-12/2 and Col-23/2, and
Ka ~ 0.3 ± 0.1 mM
1
for Col-23/3.
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Fig. 1.
Primary structures of Col-12
(A) and Col-23 (B). Numbering of
residues is as previously published (2-4). Residues of module 2 in
Col-12 and Col-23 (Col-12/2, Col-23/2) are primed (') and of module 3 in Col-23 (Col-23/3) are double-primed (") to distinguish them from
those in module 1 (Col-12/1) (unprimed). Extraneous residues stemming
from the expression vectors are shown as lowercase
letters.
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Fig. 2.
1H/15N HSQC spectra
of Col-12 (A) and Col-23 (B)
ligand-free (black) and in the presence of excess WHW
(red). The assignment of
1H/15N amide resonances has been reported
(2-4). The cross-peaks are labeled according to the residue numbering
convention described in Fig. 1. Signals from extraneous residues at the
amino terminus of Col-12 (N) have not been specifically assigned.
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Fig. 3.
Profile of WHW (A) and THS
(B) binding to Col-12 and Col-23 monitored by
NMR. The normalized resonance shifts of each FN2 module,
, corresponding to the fraction of ligand-bound
protein, are plotted versus the free ligand concentration
[L] = [Lo]
[Po]
, where
[Lo] and [Po] denote
the total ligand and total protein concentrations, respectively. The
data for Col-12/1 (blue), Col-12/2 (red),
Col-23/2 (pink), and Col-23/3 (green) are shown.
Filled symbols denote experimental data points. Each point
is an average of 3-6 selected amide 1H or 15N
normalized chemical shifts. Continuous traces represent
binding curves calculated via nonlinear least squares fit to the
experimental data.
Affinity of WHW and THS for FN2 domains
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Fig. 4.
Contact surface of Col-12/1, residues 5-59
(A), Col-12/2 (B), Col-23/2
(C), and Col-23/3, residues 3-58
(D), colored according to Col-12 or Col-23 backbone
amide chemical shift changes induced by (PPG)6
binding (2-4). Front (left) and back
(right) views are shown. The color intensity is proportional
to the sum of median-normalized amide 1H and
15N chemical shift changes of the individual residues,
scaled to achieve a balanced distribution. The atomic
coordinates on which these models are based were extracted from the
Protein Data Bank, codes 1KS0 (Col-1), 1CXW (Col-2), and 1J7M
(Col-3).
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Fig. 5.
Contact surface of Col-12/1, residues 5-59
(A), Col-12/2 (B), Col-23/2
(C), and Col-23/3, residues 3-58 (D)
colored according to Col-12 or Col-23 backbone amide chemical shift
changes induced by WHW binding. Front (left) and back
(right) views are shown. The gelatin-binding site comprising
hydrophobic cleft and a protruding loop (residues 33-38) on its
right-hand rim is revealed in the front view. The description of the
figure is the same as for Fig. 4.
View larger version (55K):
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Fig. 6.
Contact surface of Col-12/1, residues 5-59
(A), Col-12/2 (B), Col-23/2
(C), and Col-23/3, residues 3-58
(D), colored according to Col-12 or Col-23 backbone
amide chemical shift changes induced by THS binding. Front
(left) and back (right) views are shown. The
description of the figure is the same as for Figs. 4 and 5.
galCol-1 and less so
from those coated with
galCol-2 (Table I). THS perturbs amide
resonances stemming from residues on the right-hand rim of the
gelatin-binding pocket in all Col modules (Fig. 6), suggesting that THS
contacts Col modules via this site. Signals from the termini are also affected.
CONCLUSIONS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
CONCLUSIONS
REFERENCES
1 range.
Perturbation of NMR signals from the termini and the linker upon
peptide binding may reflect direct contact with the ligands or indirect
effects on conformation and dynamics of these flexible regions. The NMR
data also suggest that the contacts in most cases involve the
gelatin-binding site, particularly the protruding loop on its
right-hand rim that contains residues Gly-33 and Arg-34. The latter may
account for the competition between these peptides and gelatin for
binding to FN2 domains. Interestingly, residues Gly-33 and Arg-34 are
also involved in intramolecular interactions between the Col-3 module
and the propeptide domain (3, 5).
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ACKNOWLEDGEMENTS |
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We thank Dr. A. Patthy (Agricultural Biotechnology Center, Gödöllö, Hungary) for peptide synthesis, Professor G. P. Smith (University of Missouri, Columbia) for the generous gift of bacterial strains and phage libraries, and Dr. J. Schaller for mass spectrometry analysis of the expressed proteins.
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FOOTNOTES |
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* This work was supported by Grant CRP/HUN98-03 from the International Centre for Genetic Engineering and Biotechnology, Trieste, Italy, Grant OTKA T034317 from the Hungarian Academy of Sciences, and National Institutes of Health Grant HL29409.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.
¶ Current address: The Burnham Institute, 10901 North Torrey Pines Rd., La Jolla, CA 92037.
Current address: Dept. of Chemistry and Biochemistry,
University of Maryland, College Park, MD 20472.
** To whom correspondence should be address: Dept. of Chemistry, Carnegie Mellon University, 4400 Fifth Ave., Pittsburgh, PA 15213. E-mail: llinas@andrew.cmu.edu.
Published, JBC Papers in Press, December 16, 2002, DOI 10.1074/jbc.M210116200
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ABBREVIATIONS |
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The abbreviations used are:
MMP, matrix metalloproteinase;
FN2, fibronectin type II;
ACG, peptide
ACGYTYHPPCARLTV;
galCol-1, -2, and -3, first, second, and third FN2
modules, respectively, from human MMP-2 with amino-terminal peptide
derived from the
-galactosidase moiety of the expression vector;
galCol-123, the three in-tandem FN2 modules from human MMP-2 with
amino-terminal peptide derived from the
-galactosidase moiety of the expression vector;
Col, collagen-binding FN2 repeat;
Col-1, the
first FN2 domain from human MMP-2;
Col-12, the first and second FN2
domains from human MMP-2;
Col-12/1, the Col-1 repeat in Col-12;
Col-12/2, the Col-2 repeat in Col-12;
Col-2, the second FN2 domain from
human MMP-2;
Col-23, the second and third FN2 domains from human MMP-2;
Col-23/2, the Col-2 repeat in Col-23;
Col-23/3, the Col-3 repeat in
Col-23;
Col-3, the third FN2 domain from human MMP-2;
HAS, peptide
HASHFRFRHSHVYGV;
pro-MMP-2, the proenzyme form of MMP-2;
THS, peptide
THSHQWRHHQFPAPT;
WFP, peptide WFPGPITFIPRPWSS;
WHV, peptide
WHVSPRHQRLFHGLF;
WHW, peptide WHWRHRIPLQLAAGR.
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REFERENCES |
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---|
1. |
Collier, I. E.,
Wilhelm, S. M.,
Eisen, A. Z.,
Marmer, B. L.,
Grant, G. A.,
Seltzer, J. L.,
Kronberger, A.,
He, C. S.,
Bauer, E. A.,
and Goldberg, G. I.
(1988)
J. Biol. Chem.
263,
6579-6587 |
2. | Briknarová, K., Grishaev, A., Bányai, L., Tordai, H., Patthy, L., and Llinás, M (1999) Structure 7, 1235-1245[CrossRef][Medline] [Order article via Infotrieve] |
3. |
Briknarová, K.,
Gehrmann, M.,
Bányai, L.,
Tordai, H.,
Patthy, L.,
and Llinás, M
(2001)
J. Biol. Chem.
276,
27613-27621 |
4. | Gehrmann, M., Briknarová, K., Bányai, L., H., Patthy, L., and Llinás, M. (2002) Biol. Chem. 383, 137-148[Medline] [Order article via Infotrieve] |
5. |
Morgunova, E.,
Tuuttila, A.,
Bergmann, U.,
Isupov, M.,
Lindqvist, Y.,
Schneider, G.,
and Tryggvason, K.
(1999)
Science
284,
1667-1670 |
6. |
Collier, I. E.,
Krasnov, P. A.,
Strongin, A. Y.,
Birkedal-Hansen, H.,
and Goldberg, G. I.
(1992)
J. Biol. Chem.
267,
6776-6781 |
7. |
Tordai, H.,
and Patthy, L.
(1999)
Eur. J. Biochem.
259,
513-518 |
8. | Gehrmann, M. (2002) Structural and Functional Similarities between FII and Kringle Domains.Doctoral dissertation , Carnegie Mellon University, Pittsburgh, PA |
9. | Yu, A. E., Murphy, A. N., and Stetler-Stevenson, W. G. (1998) in Matrix Metalloproteinases (Parks, W. C. , and Mecham, R. P., eds) , pp. 85-113, Academic Press, San Diego |
10. | Itoh, T., Tanioka, M., Yoshida, H., Yoshioka, T., Nishimoto, H., and Itohara, S. (1998) Cancer Res. 58, 1048-1051[Abstract] |
11. | Koivunen, E., Arap, W., Valtanen, H., Rainisalo, A., Penate-Medina, O., Heikkilä, P., Kantor, C., Gahmber, C. G., Salo, T., Konttinen, Y. T., Sorsa, T., Rouslahti, E., and Pasqualine, R. (1999) Nat. Biotechnology 17, 768-774[CrossRef][Medline] [Order article via Infotrieve] |
12. | Talbot, D. C., and Brown, P. D. (1996) Eur. J. Cancer 32, 2528-2533[CrossRef] |
13. | Beckett, R. P., Davidson, A. H., Drummond, A. H., Huxley, P., and Whittaker, M. (1996) Drug Discov. Today 1, 16-26[CrossRef] |
14. | Santos, O., McDermott, C. D., Daniels, R. G., and Appelt, K. (1997) Clin. Exp. Metastasis 15, 499-508[CrossRef][Medline] [Order article via Infotrieve] |
15. | Davies, B., Brown, P. D., East, N., Crimmin, M. J., and Balkwill, F. R. (1993) Cancer Res. 53, 2087-2091[Abstract] |
16. | Taraboletti, G., Garofalo, A., Belotti, D., Drudis, T., Borsotti, P, Scanziani, E., Brown, P. D., and Giavazzi, R. (1995) J. Natl. Cancer Inst. 87, 293-298[Abstract] |
17. |
Volpert, O. V.,
Ward, W. F.,
Lingen, M. W.,
Chesler, L.,
Solt, D. B.,
Johnson, M. D.,
Molteni, A.,
Polverini, P. J.,
and Bouck, N. P.
(1996)
J. Clin. Invest.
98,
671-679 |
18. | Anderson, I. C., Shipp, M. A., Docherty, A. J. P., and Teicher, B. A. (1996) Cancer Res. 56, 715-718[Abstract] |
19. | Eccles, S. A., Box, G. M., Court, W. J., Bone, E. A., Thomas, W., and Brown, P. D. (1996) Cancer Res. 56, 2815-2822[Abstract] |
20. | Bányai, L., Tordai, H., and Patthy, L. (1994) Biochem. J. 298, 403-407[Medline] [Order article via Infotrieve] |
21. | Bányai, L., and Patthy, L. (1991) FEBS Lett. 282, 23-25[CrossRef][Medline] [Order article via Infotrieve] |
22. | Scott, J. K., and Smith, G. P. (1990) Science 249, 386-390[Medline] [Order article via Infotrieve] |
23. | Nishi, T., Budde, R. J. A., McMurray, J. S., Oberyesekere, N. U., Safdar, N., Levin, V. A., and Saya, H. (1996) FEBS Lett. 399, 237-240[CrossRef][Medline] [Order article via Infotrieve] |
24. | Parmley, S. F., and Smith, G. P. (1988) Gene 73, 305-318[CrossRef][Medline] [Order article via Infotrieve] |
25. | Müller, L. (1979) J. Am. Chem. Soc. 101, 4481-4484 |
26. | Bodenhausen, G., and Ruben, D. J. (1980) Chem. Phys. Lett. 69, 185-189[CrossRef] |
27. | Mori, S., Abeygunawardana, C., Johnson, M. O., and van Zijl, P. C. M. (1995) J. Magn. Reson. B 108, 94-98[CrossRef][Medline] [Order article via Infotrieve] |
28. |
Pace, C. N.,
Vajdos, F.,
Fee, L.,
Grimsley, G.,
and Gray, T.
(1995)
Protein Sci.
4,
2411-2423 |
29. | Marti, D. N., Hu, C.-K., An, S. S. A., von Haller, P., Schaller, J., and Llinás, M. (1997) Biochemistry 36, 11591-11604[CrossRef][Medline] [Order article via Infotrieve] |
30. |
An, S. S. A.,
Marti, D. N.,
Carreño, C.,
Albericio, F.,
Schaller, J.,
and Llinás, M.
(1998)
Protein Sci.
7,
1947-1959 |
31. | Ferry, G., Boutin, J. A., Atassi, G., Fauchère, J. L., and Tucker, G. C. (1996) Mol. Div. 2, 135-146 |
32. | O'Neil, K. T., Hoess, R. H., Jackson, S. A., Ramachandran, N. S., Mousa, S. A., and DeGrado, W. F. (1992) Proteins 14, 509-515[Medline] [Order article via Infotrieve] |
33. | McLafferty, M. A., Kent, R. B., Ladner, R. C., and Markland, W. (1993) Gene 128, 29-36[CrossRef][Medline] [Order article via Infotrieve] |
34. | Koivunen, E., Wang, B., and Ruoslahti, E. (1995) Bio/Technology 13, 265-270[Medline] [Order article via Infotrieve] |
35. | Foster, M. P., Wuttke, D. S., Clemens, K. R., Jahnke, W., Radhakrishnan, I., Tennant, L., Reymond, M., Chung, J., and Wright, P. E. (1998) J. Biomol. NMR 12, 51-71[CrossRef][Medline] [Order article via Infotrieve] |
36. |
Shuker, S. B.,
Hajduk, P. J.,
Meadows, R. P.,
and Fesik, S. W.
(1996)
Science
274,
1531-1534 |