MMP - Bachem

MATRIX
METALLOPROTEINASES
Matrix
Metallo
proteinases
Matrix Metalloproteinases
The matrix metalloproteinases (MMPs) constitute a multigene family
of more than 25 secreted and membrane bound enzymes that degrade
numerous pericellular substrates. Their targets include other proteinases, clotting factors, chemotactic molecules, latent growth factors, cell
surface receptors, cell-cell adhesion molecules, and almost all structural extracellular matrix proteins. Thus, MMPs play a role in normal tissue modelling and differentiation during development. In pathological
conditions they are involved in inflammation, degradation of bone, autoimmune diseases, and in the invasive migration of cancer cells across
the basement membrane as in tumor metastasis. To identify the role of
a particular MMP in these disease states, it is desirable to have a series
of synthetic peptide substrates that allow a rapid discrimination of
different MMP activities.
2
Introduction
The matrix metalloproteinases (MMPs),
also known as the ‘matrixins’, are a family of structurally and functionally related
Ca-containing Zn-endopeptidases. Together
with the serralysins, astacins, and adamalysins, they form the metzincin endopeptidase superfamily. Like all members of the
metzincins, MMPs can be distinguished by
a highly conserved motif containing three
histidines that bind zinc at the catalytic site
and a conserved methionine that is located
beneath the active site zinc. Their signature
zinc-binding motif reads HEBXHXBGBXHZ,
where histidine (H), glutamic acid (E) and
glycine (G) residues are invariant, B is a
bulky hydrophobic residue, X is a variable
residue, and Z is a family-specific amino
acid. In all but a few MMPs Z is represented
by a serine residue. MMPs are secreted as
inactive proenzymes containing a propeptide of about 80 amino acids and may be
activated by proteases or organomercurials.
Disruption of the fourth coordination site
for the catalytic zinc formed by a conserved
cysteine residue within the consensus
sequence PRCXXPD present in most MMP
propeptides is necessary for enzyme activation.
MMPs from vertebrate species are given
MMP numbers (i.e. MMP-1, MMP-2, etc.),
and MMPs from invertebrates rely on trivial
names.
MMP Structure
and Function
The MMP family currently consists of at
least 28 members, all of which share a
common catalytic core with a zinc molecule
in the active site. The MMPs can be divided
into five main subfamilies, collagenases
(MMP-1, -8, -13, and -18), stromelysins
(MMP-3, -10, and -11), gelatinases (MMP-2
and -9), matrilysins (MMP-7 and -26), and
membrane bound MMPs based on differences in structure and substrate specificity.
The membrane bound MMPs can further
be divided into two main groups, the type-I
transmembrane and the glycosylphosphatidylinositol (GPI)-anchored proteases.
MMP-14, -15, -16, and -24 are type-I trans-
membrane proteins that have a cytoplasmic
domain. The second subgroup of membrane
bound MMPs consists of enzymes which
are attached to the cell membrane by a
GPI-anchor. This subgroup includes MMP17 and MMP-25 (see Table 1). The best
characterized to date, MMP-17, has little
similarity with the other membrane-type
MMPs at the structural and functional level
and appears to be predominantly expressed
in leukocytes as well as in tumor cell lines.
All membrane-type MMPs have an arginine
rich tetrapeptide linking the propeptide and
the catalytic domain and can be activated
by furin-type convertases.
3
Matrix Metalloproteinases
Physiological
Functions of MMPs
MMPs have a wide variety of functions in
both physiological and pathological conditions. This group of proteases degrades
basal membranes and connective tissues
and plays an essential role in the homeostasis of the extracellular matrix (ECM).
Their activity is highly controlled in order to
confine them to specific areas. An activation cascade, initiated by the proteolysis of
plasminogen, cleaves proMMPs, and every
step is controlled by specific activators or
inhibitors called tissue inhibitors of metalloproteinases (TIMPs). Thus, an imbalance
in the expression or activity of MMPs can
have important consequences in diseases.
Controlled degradation of extracellular
matrix is essential in many physiological
situations including developmental tissue
remodeling, angiogenesis, tissue repair,
and normal turnover of ECM (the female
reproductive cycle, bone remodeling,
nerve growth, hair follicle cycling, immune
response, and apoptosis). However, their
unregulated activity has been implicated
in numerous disease processes including
arthritis (there is uncontrolled destruction
of cartilage), multiple sclerosis, cardiovas-
Collagenases
Stromelysins
Gelatinases
Matrilysins
Membrane bound MMPs
Type-I transmembrane
GPI-anchored
4
cular diseases, neurological diseases, blood
brain barrier damage, nephritis, corneal
ulceration, skin ulceration, gastric ulcer,
liver cirrhosis, fibrotic lung disease, vascular diseases, Alzheimer´s disease, infertility,
and tumor cell metastasis.
Therefore, the characterization of MMPs
involved in specific types and stages
of tumors will significantly improve the
diagnosis and treatment of these cancers
in humans. As there is growing evidence
that malignant tumors use MMPs for tumor
growth and spreading, low molecular weight
inhibitors of these proteins have encouraged structural studies on several members
of the MMP family, so that the molecular
details of enzyme-inhibitor interactions of
the MMPs have become available.
In this context, synthetic MMP inhibitors
and substrates should lead to the development of a new generation of anticancer
agents with additional beneficial properties
compared to the existing cytotoxic agents
used in the treatment of human malignancies.
MMP-1, -8, -13, and -18
MMP-3, -10, and -11
MMP-2 and -9
MMP-7 and -26
MMP-14, -15, -16, and -24
MMP-17 and -25
Table 1.
MMP Subfamilies
Table 2.
The Vertebrate Matrix Metalloproteinases
MMP
Common Name
MMP-1
Mol. Wt.
(latent/active)
E.C.No.
MEROPS ID
Substrate
Interstitial collagenase-1, Col- 55000/45000
lagenase-1
3.4.24.7
M10.001
CN types I, II, III, V, VII, VIII, and X,
aggrecan, gelatin, serpins
MMP-2
Gelatinase A, 72 kD Gelatinase, Type IV Collagenase
72000/66000
3.4.24.24
M10.003
CN types I, IV, V, VII, and X, gelatin,
elastin, FN, LN, MMP-9, MMP-13
MMP-3
Stromelysin-1, Proteoglycanase, Transin, Procollagenase
57000/45000
3.4.24.17
M10.005
CN types II, IV, IX, X, and XI, LN, FN,
proteoglycan, aggrecan, elastin,
gelatin, proMMP-1, proMMP-8,
proMMP-9
MMP-7
Matrilysin, Matrin, Uterine
Metalloendopeptidase, Putative Metalloproteinase1 (PUMP-1)
28000/19000
3.4.24.23
M10.008
CN type IV, elastin, proteoglycan,
glycoprotein, gelatin
MMP-8
Neutrophil Collagenase, Collagenase-2
75000/58000
3.4.24.34
M10.002
CN types I, II, III, and V
MMP-9
Gelatinase B, Type V Collagenase, 92 kD Type IV Collagenase, Macrophage Gelatinase
92000/86000
3.4.24.35
M10.004
CN types IV, gelatin, aggrecan, LN
MMP-10
Stromelysin-2, Transin-2
57000/44000
3.4.24.22
M10.006
CN types I, II, III, and V
MMP-11
Stromelysin-3
51000/44000
3.4.24.B3
M10.007
LN, α1-proteinase inhibitor, α1antitrypsin
MMP-12
Macrophage Elastase, MME,
Metalloelastase
54000/45000 and 3.4.24.65
22000
M10.009
Elastin
MMP-13
Collagenase-3
60000/48000
3.4.24.B4
M10.013
CN types I, II, III, IV, V, IX, X, and XI,
gelatin, LN, tenascin, aggrecan, FN
MMP-14
MT1-MMP, Membrane-type
Matrix Metalloproteinase-1
66000/56000
3.4.24.80
M10.014
CN types I, II, and III, gelatin,
FN, LN, VN, aggrecan, tenascin,
perlecan, proMMP-2, proMMP-13,
proteoglycan
MMP-17
MT4-MMP, Membrane-type
Matrix Metalloproteinase-4
57000/53000
3.4.24.-
M10.017
FN, fibrin, gelatin
MMP-25
MT6-MMP, Leukolysin,
Membrane-type Matrix Metalloproteinase-6
3.4.24.-
M10.024
proMMP-2
MMP-26
Endometase, Matrilysin-2
3.4.24.B7
M10.029
CN type IV, FN, FG
MMP-28
Epilysin
3.4.24.-
unknown
Abbreviations: CN = collagen; FN = fibronectin; LN = laminin; VN = vitronectin; FG = fibrinogen
5
Matrix Metalloproteinases
Cancer cell,
SEM
Cancer cell.
Coloured scanning electron micrograph
(SEM) of a spreading cancer cell, showing its uneven surface and cytoplasmic
projections. Clumps of cancerous cells
form tumours, which possess the ability to invade and destroy surrounding
tissues and travel to distant parts of
the body to seed secondary tumours.
Malignant cells proliferate and grow in a
chaotic manner, with defective cell division retained within each new generation of cells. Variations also occur in the
size and structure of the cells. The cell
here is taken from a breast cancer.
KEYSTONE/SCIENCE PHOTO LIBRARY/
NATIONAL CANCER INSTITUTE
6
references
N. Borkakoti
Matrix metalloproteases: variations
on a theme.
Prog. Biophys. Mol. Biol. 70, 73-94
(1998)
W.C. Parks
Matrix metalloproteinases in repair.
Wound Repair Regen. 7, 423-432
(1999)
M.A. Forget et al.
Physiological roles of matrix metalloproteinases: implications for
tumor growth and metastasis.
Can. J. Physiol. Pharmacol. 77, 465480 (1999)
N. Borkakoti
Structural studies of matrix metalloproteinases.
J. Mol. Med. 78, 261-268 (2000)
P.Z. Khasigov et al.
Matrix metalloproteinases of normal
human tissues.
Biochemistry 66,130-140 (2001)
G. Cox and K.J. O‘Byrne
Matrix metalloproteinases and
cancer.
Anticancer Res. 21, 4207-4219
(2001)
V. Ellis and G. Murphy
Cellular strategies for proteolytic
targeting during migration and invasion.
FEBS Lett. 506,1-5 (2001)
M.D. Sternlicht and Z. Werb
How matrix metalloproteinases
regulate cell behavior.
Annu. Rev. Cell Dev. Biol. 17, 463-516
(2001)
D.G. Armstrong and E.B. Jude
The role of matrix metalloproteinases in wound healing.
J. Am. Podiatr. Med. Assoc. 92, 12-18
(2002)
L.M. Coussens et al.
Matrix metalloproteinase inhibitors
and cancer: trials and tribulations.
Science 295, 2387-2392 (2002)
Y. Jiang et al.
Complex roles of tissue inhibitors of
metalloproteinases in cancer.
Oncogene 21, 2245-2252 (2002)
Z.S. Galis and J.J. Khatri
Matrix metalloproteinases in vascular remodeling and atherogenesis:
the good, the bad, and the ugly.
Circ. Res. 90, 251-262 (2002)
M.V. Cronauer et al.
Fibroblast growth factors and their
receptors in urological cancers:
basic research and clinical implications.
Eur. Urol. 43, 309-319 (2003)
U. Ikeda and K. Shimada
Matrix metalloproteinases and coronary artery diseases.
Clin. Cardiol. 26, 55-59 (2003)
A. Mancini and J.A. Di Battista
Transcriptional regulation of matrix
metalloprotease gene expression in
health and disease.
Front. Biosci. 11, 423-446 (2006)
E.I. Deryugina and J.P. Quigley
Matrix metalloproteinases and
tumor metastasis.
Cancer Metastasis Rev. 25, 9-34
(2006)
7
MMP
Substrates
and
Inhibitors
Matrix Metalloproteinases
Additionally to our comprehensive choice of MMP substrates and inhibitors presented in this brochure, we offer a host of chromogenic, fluorogenic, and FRET substrates for proteases and other enzymes. Our brochure
FRET Substrates, which can be downloaded from our homepage gives you
an overview on these types of substrates.
Our website offers further publications presenting products for enzyme
research, Secretase Substrates Inhibitors and Caspase Substrates Inhibitors, for download.
If you are looking for a substrate or an inhibitor for your enzyme in our
online shop, the search could be facilitated by using the Inhibitors & Substrates index:
 Inhibitors & Substrates
 Choose the enzyme
8
MMP-1
Substrates
Ac-Pro-Leu-Gly-[(S)-2-mercapto4-methyl-pentanoyl]-Leu-Gly-OEt
H-7145
Ac-PLG-psi[COS]LLG-OEt
Dnp-Pro-Leu-Gly-Cys(Me)-His-Ala-DArg-NH2
M-1905
Dnp-PLGC(Me)HAr-NH2
Dnp-Pro-β-cyclohexyl-Ala-AbuCys(Me)-His-Ala-Lys(N-Me-Abz)-NH2
M-1910
Dnp-P-Cha-Abu-C(Me)HAK(N-Me-Abz)NH2
FA-Leu-Gly-Pro-Ala-OH
M-1385
FA-LGPA
Dnp-Pro-Gln-Gly-OH
M-1335
Dnp-PQG
Dnp-Pro-Gln-Gly-Ile-Ala-Gly-Gln-DArg-OH
M-1340
Dnp-PQGIAGQr
MMP-1
Inhibitors
Z-Gly-Pro-Gly-Gly-Pro-Ala-OH
M-1260
Z-GPGGPA
Z-Gly-Pro-Leu-Gly-Pro-OH
M-1265
Z-GPLGP
4-Abz-Gly-Pro-D-Leu-D-Ala-NHOH
N-1405
4-Abz-GPla-NHOH
Z-Pro-Leu-Ala-NHOH
N-1285
Z-PLA-NHOH
H-Cys-OH · HCl · H2O
E-1755
Z-Pro-D-Leu-D-Ala-NHOH
N-1290
Z-Pla-NHOH
1,4-Dithio-DL-threitol
Q-1225
Phenylmethylsulfonyl fluoride
Q-1395
MMP-2
Substrates
Pz-Pro-Leu-Gly-Pro-D-Arg-OH
M-1715
Pz-PLGPr
DABCYL-γ-Abu-Pro-Gln-Gly-LeuGlu(EDANS)-Ala-Lys-NH2
M-2495
DABCYL-γ-Abu-PQGLE(EDANS)AK-NH2
Mca-(Ala7,Lys(Dnp)9)-Bradykinin
M-2405
Mca-RPPGFSAFK(Dnp)
Z-Pro-Leu-Gly-NHOH
C-3205
Z-PLG-NHOH
Mca-Pro-Leu-OH
M-1975
Mca-PL
Amyloid β-Protein (10-20)
H-1388
YEVHHQKLVFF
Mca-Pro-Leu-Gly-Leu-Dap(Dnp)-AlaArg-NH2
M-1895
Mca-PLGL-Dpa-AR-NH2
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Matrix Metalloproteinases
MMP-2
Inhibitors
H-Cys-OH · HCl · H2O
E-1755
1,4-Dithio-DL-threitol
Q-1225
H-Cys-Thr-Thr-His-Trp-Gly-Phe-ThrLeu-Cys-OH
H-4736
CTTHWGFTLC
H-Gly-OH
E-3000
Decanoyl-Arg-Val-Lys-Arg-chloromethylketone
N-1505
Dec-RVKR-CMK
Peptide 74
H-8545
TMRKPRCGNPDVAN
MMP-3
Substrates
DABCYL-γ-Abu-Arg-Pro-Lys-Pro-ValGlu-Nva-Trp-Arg-Glu(EDANS)-Ala-LysNH2
M-2490
DABCYL-γ-Abu-RPKPVE-NvaWRE(EDANS)AK-NH2
6-(7-Nitro-benzo[2,1,3]oxadiazol4-ylamino)-hexanoyl-Arg-Pro-LysPro-Leu-Ala-Nva-Trp-Lys((7-dimethylaminocoumarin-4-yl)-acetyl)-NH2
M-2300
NBD-ε-aminocaproyl-RPKPLA-NvaWK(DMACA)-NH2
MMP-3
Inhibitors
Acetyl-Stromelysin-1 Precursor (9196) amide (human, horse, mouse)
H-2504
Ac-RCGVPD-NH2
H-Cys-OH · HCl · H2O
E-1755
MMP-7
Substrates
Dnp-Arg-Pro-Leu-Ala-Leu-Trp-ArgSer-OH
M-2205
Dnp-RPLALWRS
MMP-8
Inhibitors
H-Pro-Leu-Gly-NHOH · HCl
M-1775
PLG-NHOH · HCl
10
MMP-9
Substrates
Dnp-Pro-β-cyclohexyl-Ala-GlyCys(Me)-His-Ala-Lys(N-Me-Abz)-NH2
M-2055
Dnp-P-Cha-GC(Me)HAK(N-Me-Abz)NH2
Dnp-Pro-Leu-Gly-Leu-Trp-Ala-D-ArgNH2
M-1855
Dnp-PLGLWAr-NH2
MMP-12
Substrates
Mca-Arg-Pro-Lys-Pro-Gln-OH
M-2250
Mca-RPKPQ
Mca-Pro-Leu-Gly-Leu-Glu-Glu-AlaDap(Dnp)-NH2
M-2670
Mca-PLGLEEA-Dpa-NH2
Mca-Arg-Pro-Lys-Pro-Tyr-Ala-NvaTrp-Met-Lys(Dnp)-NH2
M-2105
Mca-RPKPYA-Nva-WMK(Dnp)-NH2
Mca-Arg-Pro-Lys-Pro-Val-Glu-NvaTrp-Arg-Lys(Dnp)-NH2
M-2110
Mca-RPKPVE-Nva-WRK(Dnp)-NH2
MMP-13
Substrates
Dnp-(Leu421)-Collagen Type VIII α1
Chain (419-426) amide (human, mouse)
M-2530
Dnp-GPLGMRGL-NH2
Mca-Pro-Lys-Pro-Leu-Ala-LeuDap(Dnp)-Ala-Arg-NH2
M-2225
Mca-PKPLAL-Dpa-AR-NH2
Mca-(endo-1a-Dap(Dnp))-TNF-α (-5 to
+6) amide (human)
M-2255
Mca-PLAQAV-Dpa-RSSSR-NH2
Mca-Pro-β-cyclohexyl-Ala-Gly-NvaHis-Ala-Dap(Dnp)-NH2
M-2515
Mca-P-Cha-G-Nva-HA-Dpa-NH2
Mca-Lys-Pro-Leu-Gly-Leu-Dap(Dnp)Ala-Arg-NH2
M-2350
Mca-KPLGL-Dpa-AR-NH2
MMP-14
Substrates
Mca-Pro-Leu-Ala-Cys(Mob)-Trp-AlaArg-Dap(Dnp)-NH2
M-2510
Mca-PLAC(Mob)WAR-Dpa-NH2
MMP-26
Substrates
Mca-Pro-Leu-Ala-Nva-Dap(Dnp)-AlaArg-NH2
M-2520
Mca-PLA-Nva-Dpa-AR-NH2
11
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Published by Global Marketing, Bachem Group, March 2014
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