Matrix Metalloproteinases (MMPs)

First printed in R&D Systems' 1999 Catalog.



The MMP family of enzymes contributes to both normal and pathological tissue remodeling. MMPs play a key role in the migration of normal and malignant cells through the body. They also act as regulatory molecules, both by functioning in enzyme cascades and by processing matrix proteins, cytokines, growth factors and adhesion molecules to generate fragments with enhanced or reduced biological effects.

Domain Structure & Function

Figure 1. MMPs can facilitate tumor cell metastasis and angiogenesis. Adapted from Opdenakker, G. & J. Van Damme (1992) Cytokine 4:251.

The matrix metalloproteinases (MMPs) are members of a family of at least 15 Zn-dependent endopeptidases that function extracellularly (Table 1).1 The MMPs each contain a protease domain that has a conserved HExGHxxGxxHS/T sequence in which the three Histidine residues form a complex with a catalytic Zn atom. In addition, all MMPs contain a regulatory domain (pro-piece) with a conserved PRCGxPD motif that is responsible for maintaining latency in MMPs via binding of the cysteine residue to the active site Zn. The simplest MMP is MMP-7 (matrilysin), which consists of a pro-piece and catalytic domain only. The other MMPs maintain this basic unit but have a variable number of structural domains added. Although most MMPs are secreted proteins, the recently described membrane-type MMPs (MT-MMPs) are anchored to the cell membrane by a transmembrane and intracytoplasmic domain. X-ray crystallography has shown that the catalytic domains of the different MMPs have similar structure, but the topology of the active site clefts differs, accounting for some of the differences in substrate specificities. Differences in the other domains confers further substrate specificity, regulates binding to matrix proteins, and determines interactions with the Tissue Inhibitors of Metalloproteinases (TIMPs), the natural inhibitors of MMP activity.2

Substrates & Nomenclature

Much of the early literature suggested that each MMP had its own particular substrate.1 This concept led to the use of substrate-focused nomenclature for MMPs such that the collagenases broke down intact fibrillar collagens, gelatinases degraded denatured collagen, and metalloelastase attacked elastin. It is now recognized that MMPs usually degrade multiple substrates, with considerable substrate overlap between individual MMPs. For example, interstitial collagenase (MMP-1) is capable of degrading casein, gelatin, a-1 antitrypsin, myelin basic protein, L-Selectin, pro-TNF and IL-1 beta and pro-MMP-2 and -9. 72-kDa gelatinase (MMP-2) can degrade fibrillar collagen, elastin, IGF-binding proteins, FGF receptor and can activate MMP-1, -9 and -13. MMP-12 is highly active against type IV collagen, gelatin, fibronectin, vitronectin and plasminogen, but it is not very effective at degrading elastin. See Table 1 for a list of substrates that can be cleaved by purified MMPs in vitro.

In an attempt to break the link between name and function, all MMPs are now given an MMP number, such that Interstitial collagenase is MMP-1, etc. (Table 1). There are holes in this system. There is no MMP-4, -5 or -6, as the activities could not be ascribed to a specific gene product, and MMP-18 is known only as a Xenopus enzyme. As with all other enzymes, MMPs have an EC classification, although this lags well behind the MMP designation.

Table 1. MMP Facts.
MMP Alternative Names EC Number Chromosome Substrates
  • Collagenase
  • Fibroblast Collagenase
  • Interstitial Collagenase
EC3.4.24.7 11q22-q23 Collagens (I, II, III, VII, VIII and X)17, 18; Gelatin; aggrecan19; hyaluronidase-treated versican20; proteoglycan link protein21; large tenascin-C22; a1-antitrypsin/a1-proteinase inhibitor (a1-AT)23-25; a1-antichymotrypsin (a1-ACHYM)24, 25; a2M; rat a1M; pregnancy zone protein; rat a1I3 (a1-inhibitor 3); ovostatin; entactin (nidogen)26; MBP27; GST-TNF/TNF peptide28; L-Selectin29; IL-1ß30; serum amyloid A; IGF-BP532; IGF-BP333; MMP-234; MMP-920
  • 72 kDa Gelatinase
  • Gelatinase A
  • Type IV Collagenase
  • Neutrophil Gelatinase
EC3.4.24.24 16q13 Collagens (I49, IV50, V, VII, X17, XI and XIV51); Gelatin; elastin50, 52; fibronectin; laminin-1, laminin-553; galectin-354; aggrecan45; decorin55; hyaluronidase-treated versican20; proteoglycan link protein21; osteonectin48; MBP27; GST-TNF/TNF peptide28; IL-1ß56; Aß1-4056, 57v; Ab10-2058; APP69559; a1-AT25; prolysyl oxidase fusion protein60; IGF-BP532; IGF-BP333; FGF R161; MMP-134; MMP-962; MMP-1363
  • Stromelysin-1
  • Transin
EC3.4.24.17 11q23 Collagens (III, IV50, V, IX); Gelatin; aggrecan19, 66; versican and hyaluronidase-treated versican20; perlecan46; decorin55; proteoglycan link protein21; large tenascin-C23; fibronectin; laminin; entactin26, 67; osteonectin48; elastin40; casein68; a1-AT23, 25; a1-ACHYM25; antithrombin-III2525; a2M; ovostatin; Substance P; MBP27; GST-TNF/TNF peptide28; IL-1ß30; serum amyloid A31; IGF-binding protein-333; fibrinogen and cross-linked fibrin69; plasminogen70; MMP-1 “superactivation”71; MMP-2/TIMP-2 complex72; MMP-773; MMP-874; MMP-975; MMP-1343
  • Matrilysin
  • PUMP
EC3.4.24.23 11q21-q22 Collagen IV82, 50 and X18; Gelatin82; aggrecan19; decorin55; proteoglycan link protein21; fibronectin and laminin82; insoluble fibronectin fibrils83; entactin26; large and small tenascin-C84; osteonectin48; ß4 intergrin85; elastin50; casein82; transferrin86; MBP27; a1-AT23; GST-TNF/TNF peptide28; plasminogen65; MMP-173; MMP-287; MMP-973, 35, 88; MMP-9/TIMP-1 complex88
  • Neutrophil Collagenase
  • Collagenase I
EC3.4.24.34 11q21-q22 Collagens (I, II, III, V36, VII, VIII and X); Gelatin; aggrecan37, 38; a1-AT39; a1-ACHYM16; a2-antiplasmin40; fibronectin41
  • 92 kDa Gelatinase
  • Gelatinase B
EC3.4.24.35 20q11.2-q13.1 Collagens (IV50, V36, VII, X17 and XIV51); Gelatin; elastin50, 52; galectin-354; aggrecan19; hyaluronidase-treated versican20; proteoglycan link protein21; fibronectin31; entactin26; osteonectin48; a1-AT23, 24; MBP27; GST-TNF/TNF peptide28; IL-1ß30; Aß1-4064; plasminogen65
  • Stromelysin-2
EC3.4.24.22 11q22.3-q23 Collagens (III76, IV77, 50 and V76); Gelatin74; casein68, 76; aggrecan66; elastin50; proteoglycan link protein21; MMP-168; MMP-877
  • Stromelysin-3
  22q11.2 Human enzyme: a1-AT78; a2M79; casein79; IGF-binding protein-180; Mouse enzyme: a1-AT78; casein, laminin, fibronectin, gelatin, collagen IV and carboxymethylated transferrin81
  • Macrophage Metalloelastase
  11q22.2-q22.3 Collagen IV89; Gelatin89; elastin and k-elastin60, 90; casein91; a1-AT89, 92; fibronectin89; vitronectin89; laminin89; entactin93; proteoglycan monomer89, 77, 94; GST-TNF89; MBP89; fibrinogen94; fibrin95; plasminogen96
  • Collagenase-3
  11q22.3 Collagens (I, II and III42, 43, IV, IX, X and XIV44); Gelatin, a1-ACHYM and plasminogen activator inhibitor 244; aggrecan45; perlecan46; large tenascin-C and fibronectin44; osteonectin47; MMP-948
  • MT-MMP-1
  14q11-q12 Collagen (I, II and III97, 98); Gelatin, casein, k-elastin, fibronectin, laminin, vitronectin and proteoglycans97-99; large tenascin-C, entactin98; a1-AT, a2M97; GST-TNF98; MMP-2100, 101; MMP-1363
  • MT-MMP-2
  16q12.2-q21 Fibronectin, large tenascin-C, entactin, laminin, aggrecan, perlecan98; GST-TNF98; MMP-2102, 103
  • MT-MMP-3
  8q21 Collagen III104; Gelatin, casein105; fibronectin; MMP-2102, 103
  • MT-MMP-4
MMP-19<     12q14 Gelatin107*, 108
  • Enamelysin
  ND Amelogenin109
* assigned as MMP-18 in reference 107. MMP-18 has, however, subsequently been assigned as xenopus MMP

Role in Physiology and Pathology

Although the link between single MMPs and individual substrates is not as direct as once thought, it is clear that as a family, the MMPs are capable of breaking down any extracellular matrix component (see Table 1). In normal physiology, MMPs produced by connective tissue are thought to contribute to tissue remodeling in development, in the menstrual cycle, and as part of repair processes following tissue damage. The obvious destructive capability of MMPs initially focused most research onto diseases that involve breakdown of the connective tissues (e.g., rheumatoid arthritis, cancer and periodontal disease). Leukocytes, particularly macrophages, are major sources of MMP production. MMPs released by leukocytes play vital roles in allowing leukocytes to extravasate and penetrate tissues, a key event in inflammatory disease. Opdenakker proposed that MMP action not only permits leukocyte emigration into tissues and causes tissue damage, it also generates immunogenic fragments of normal proteins that may escalate autoimmune disease. In an analogous way, metastatic cancer cells also use MMPs to get in and out of tissues and to establish a blood supply (Figure 1).4 Drug companies have synthesized low molecular weight MMP inhibitors that have shown efficacy in models of these diseases, reinforcing their central role in pathology.5

The MMP axis is highly regulated to avoid excessive tissue damage. Most MMPs, with the exception of 72 kDa gelatinase and the MT-MMPs, are not constitutively expressed in normal tissues. Inflammatory cytokines such as IL-1 and TNF, growth factors such as TGF-beta and noxious stimuli are required to initiate transcription. MMPs are also expressed as inactive zymogens (the pro-piece must be dissociated from the catalytic domain before the enzyme is activated). This dissociation can be achieved by autocatalysis or by the action of enzymes such as furin, plasmin or even other MMPs. For example, the activation of pro-MMP-2 occurs at the surface of many cells and is mediated by MT-MMPs. Once activated, MMPs are subject to inactivation by TIMPs and by binding to plasma proteins such as alpha-2 macroglobulin. It is thought that the local balance of MMP expression and activation versus the level of TIMP governs the level of destruction mediated by MMPs. This is of great significance when studying MMP involvement in disease processes.

In order to implicate a particular MMP in a disease, several overlapping approaches have been taken. Each has its advantages and disadvantages (Table 2). Development of a comprehensive picture of MMP involvement in any tissue culture system or in vivo disease model likely would require several of these methods.

Table 2. Methods for studying the involvement of MMPs in desease
Quantitative PCR
  • MMP specific
  • Can measure changes in mRNA for multiple MMPs in small samples
  • High throughput
  • Primers easy to design and test
  • Does not measure protein or activity
  • Can be MMP specific
  • Can localize MMP expression
  • Antibodies can cross react with other MMPs
  • Does not discriminate active and inactive enzyme
  • Labor intensive
  • Low sensitivity
  • MMP specific
  • High throughput of samples
  • MMPs found in blood and tissue fluids
  • High sensitivity possible
  • May crossreact with other MMPs
  • May detect inactive or inhibitor -complexed MMPs
Activity assays
  • Can detect active MMP
  • Can be very sensitive
  • May not be specific for particular MMP
  • Subject to sample interference
Gene knock out in mice
  • MMP specific
  • Costly
  • Requires establishment of disease model in knock out strain
  • Knock out may be compensated for in development
MMP inhibitors in models
  • Direct relevance to therapy
  • MMP specific inhibitors have not yet been described



The MMP/Cytokine Connection



Figure 2. The MMP/Cytokine Connection

The MMP axis has several areas of overlap with the cytokine network. As described above, inflammatory cytokines or growth factors can regulate the expression of MMPs. Cytokine activation of cells can also lead to increased processing of MMPs from the inactive zymogens to the active enzymes. Cytokines and their receptors can also be substrates for MMP action (Figure 2). Pro-inflammatory cytokine IL-1 beta can be cleaved and inactivated by MMP-1, -2, -3, and -9.9 In addition, the degradation of matrix proteins such as decorin can liberate growth factors such as TGF-beta that are sequestered on the matrix.10 Many membrane-bound cytokines, receptors and adhesion molecules can be released from the cell surface by the action of metalloproteinases, referred to as sheddases or convertases.11, 12 This may down regulate cell surface signaling by removal of a receptor or extend the sphere of influence of a molecule by release of a soluble active form. The consequences of this will depend on the molecule. For example, soluble TNF cleaved from the cell surface is pro-inflammatory, whereas TNF receptors cleaved from a cell act as soluble TNF inhibitors. In contrast, the cleaved soluble IL-6 receptor acts to stabilize IL-6 and the complex acts as an IL-6 agonist.

Although classical MMPs can process many of these cell surface molecules, members of the reprolysin or adamalysin clan of metalloenzymes may contribute much of the sheddase or convertase activity at the cell membrane. These enzymes have a catalytic site similar to that of the MMPs, but they have a different domain structure. The best characterized enzyme is the TNF-alpha converting enzyme (TACE or ADAM-17).13-14 This enzyme was initially isolated from cell membranes using a TNF substrate assay to follow purification. Combined inhibitors of MMP and sheddase activity have been produced. They are active in models of inflammatory diseases, such as multiple sclerosis, where a cytokine drive has been implicated.15


  1. Nagase, H. (1996) Matrix Metalloproteinases in Zinc Metalloproteases in Health and Disease, Hooper NM, ed., Taylor and Francis, London.
  2. Gomez, D.E. et al. (1997) Eur. J. Cell Biol. 74:111.
  3. Rawlings, N.D. & A.J. Barrett (1995) Methods in Enzymology 348:183.
  4. Opdenakker, G & J. Van Damme (1992) Cytokine 4:251.
  5. Beckett, R.P. et al. (1996)Drug Development Today 1:16.
  6. Hughes, P.M. et al. (1998) Brain 121:481.
  7. Maeda, A. & R.A. Sobel (1996) J. Neuropathol. Exp. Neurol. 55:300.
  8. Brown, P.D. et al. (1993) J. Natl. Cancer Inst. 85:574.
  9. Ito, A. et al. (1996) J. Biol. Chem. 271:14657.
  10. Imai, K. et al. (1997) J. Biol. Chem 322:809.
  11. Arribas, J. et al. (1996) J. Biol. Chem. 271:11376.
  12. Hooper, N.M. et al. (1997) Biochem J. 321:265.
  13. Black, R. et al. (1997) Nature 385:729.
  14. Moss M.L. et al. (1997) Nature 385:733.
  15. Chandler, S. et al. (1997) J. Neuroimmunol. 72:155.
  16. Birkedal-Hansen, H. et al. (1993) Crit. Rev. Oral Biol. Med. 4:197.
  17. Welgus, H. et al. (1990) J. Biol. Chem. 265:13521.
  18. Sires, U.I. et al. (1995) J. Clin. Invest. 95:2089.
  19. Fosang, A.J. et al. (1993) Biochem. J. 295:273.
  20. Perides, G. et al. (1995) Biochem. J. 312:377.
  21. Nguyen, Q. et al. (1993) Biochem. J. 295:595.
  22. Imai, K. et al. (1994) FEBS Lett. 352:216.
  23. Sires, U.I. et al. (1994) Biochem. Biophys. Res. Commun. 204:613.
  24. Desrochers, D.E. et al. (1991) J. Clin. Invest. 87:2258.
  25. Mast, A. et al. (1991) J. Biol. Chem. 266:15810.
  26. Sires, U.I. et al. (1993) J. Biol. Chem. 268:2069.
  27. Chandler, S. et al. (1995) Neurosci. Lett. 201:223.
  28. Gearing, A.J.H. et al. (1994) Nature 370:555.
  29. Preece, G. et al. (1996) J. Biol. Chem. 271:11634.
  30. Ito, A. et al. (1996) J. Biol. Chem. 271:14657.
  31. Mitchell, T.I. et al. (1993) Biochim. Biophys. Acta. 1156:245.
  32. Thrailkill, K.M. et al. (1995) Endocrinology 136:3527.
  33. Fowlkes, J.L. et al. (1994) J. Biol. Chem. 269:25742-25746.
  34. Crabbe, T. et al. (1994) Biochem. 33:14419.
  35. Sang, Q-X. et al. (1995) Biochim. Biophys. Acta. 1251:99.
  36. Murphy, G. et al. (1982) Biochem. J. 203:209.
  37. Fosang, A.J. (1994) Biochem. J. 304:347.
  38. Fosang, A. et al. (1992) J. Biol. Chem. 267:19470.
  39. Desrochers, P.E. et al. (1992) J. Biol. Chem. 267:5005.
  40. Knäuper, V. et al. (1993) Biochem. J. 291:847.
  41. Tschesche, H. et al. (1992) Matrix Suppl. No. 1, 245.
  42. Freije, J.M.P. et al. (1994) J. Biol. Chem. 269:16766.
  43. Knäuper, V. et al. (1996) J. Biol. Chem. 271:1544.
  44. Knäuper, V. et al. (1997) J. Biol. Chem., 272:7608.
  45. Fosang, A.J. et al. (1996) FEBS Lett. 380:17.
  46. Whitelock, J. M. et al. (1996) J. Biol. Chem. 271:10079.
  47. Knäuper, V. et al. (1997) Eur. J. Biochem. 248:369.
  48. Sasaki, T. et al. (1997) J. Biol. Chem. 272:9237.
  49. Aimes, R. and J. Quigley (1995) J. Biol. Chem. 270:5872.
  50. Murphy, G. et al. (1991) Biochem. J. 277:277.
  51. Sires, U.I. et al. (1995) Biol. Chem. 270:1062.
  52. Senior, R.M. et al. (1991) J. Biol. Chem. 266:7870.
  53. Giannelli, G. et al. (1997) Science 277:225.
  54. Ochieng, J. et al. (1994) Biochem. 33:14109.
  55. Imai, K. et al. (1997) Biochem. J. 322:809.
  56. Roher, A.E. et al. (1994) Biochem. Biophys. Res. Commun. 205:1755.
  57. Walsh, D.M. et al. (1994) Nature 367:27.
  58. Miyazaki, K. et al. (1993) Nature 362:839.
  59. LePage, R.N. et al. (1995) FEBS Lett. 377:267.
  60. Panchenko, M.V. et al. (1996) J. Biol. Chem. 271:7113.
  61. Levi, E. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7069.
  62. Fridman, R. et al. (1995) Cancer Res. 55:2548.
  63. Knäuper, V. et al. (1996) J. Biol. Chem. 271:17124.
  64. Backstrom, J.R. et al. (1996) J. Neurosci. 16:7910.
  65. Patterson, B.C. and Q.X.A. Sang (1997) J. Biol. Chem. 272:28823.
  66. Fosang, A.J. et al. (1991) J. Biol. Chem. 266:15579.
  67. Alexander, C.M. et al. (1996) J. Cell Biol. 135:1669.
  68. Windsor, L.J. et al. (1993) J. Biol. Chem. 268:17341.
  69. Bini, A. et al. (1996) Biochem. 35:13056.
  70. Lijnen, H.R. et al. (1998) Biochem. 37:4699.
  71. Murphy, G. et al. (1987) Biochem. J. 248:265.
  72. Miyazaki, K. et al. (1992) Biochem. Biophys. Res. Comm. 185:852.
  73. Imai, K. et al. (1995) J. Biol. Chem., 270:6691-6697. See also (17) above.
  74. Knäuper, V. et al. (1993) Biochem. J. 295:581.
  75. Shapiro, S.D. et al. (1995) J. Biol. Chem., 270:6351.
  76. Nicholson, R. et al. (1989) Biochem. 28:5195.
  77. Knäuper, V. et al. (1995) Int. J. Exp. Pathol. 76:A19; Knäuper, V. et al. (1996) J. Biol. Chem. 235:187.
  78. Noël, A. et al. (1995) J. Biol. Chem. 270:22866.
  79. Pei, D. et al. (1994) J. Biol. Chem. 269:25849.
  80. Ma_es, S. et al. (1997) J. Biol. Chem. 272:25706.
  81. Murphy, G. et al. (1993) J. Biol. Chem. 268:15435.
  82. Miyazaki, K. et al. (1990) Cancer Res. 50:7758.
  83. von Bredow, D.C. et al. (1995) Exp. Cell Res. 221:83.
  84. Siri, A. et al. (1995) J. Biol. Chem. 270:8650.
  85. von Bredow, D.C. et al. (1997) Exp. Cell Res. 236:341.
  86. Abramson, S.R. et al. (1995) J. Biol. Chem. 270:16016.
  87. Crabbe, T. et al. (1994) FEBS Lett. 345:14.
  88. von Bredow, D.C. et al. (1998) Biochem. J. 331:965.
  89. Chandler, S. et al. (1996) Biochem. Biophys. Res. Commun. 228:421.
  90. Shapiro, S.D. et al. (1993) J. Biol. Chem. 268:23824.
  91. Shipley, J.M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:3942.
  92. Banda, M.J. et al. (1987) J. Clin. Invest. 79:1314.
  93. Gronski, T.J. et al. (1997) J. Biol. Chem. 272:12189.
  94. Banda, M.J. and Z. Werb (1980) Fed. Proc. Fed. Am. Soc. Exp. Biol. 39:1756.
  95. Banda, M.J. and Z. Werb (1979) Fed. Proc. Fed. Am. Soc. Exp. Biol. 38:1339.
  96. Dong, Z. et al. (1997) Cell 88:801.
  97. Ohuchi, E. et al. (1997) J. Biol. Chem. 272:2446.
  98. d'Ortho, M-P. (1997) Eur. J. Biochem. 250:751.
  99. Pei, D. and S. Weiss (1996) J. Biol. Chem. 271:9135.
  100. Sato, H. et al. (1994) Nature 370:61.
  101. Will, H. et al. (1996) J. Biol Chem. 271:17119.
  102. Tanaka, M. et al. (1997) FEBS Lett. 402:219.
  103. Butler, G.S. et al. (1997) Eur. J. Biochem. 244:653.
  104. Matsumoto, S. et al. (1997) Biochim. Biophys. Acta 1354:159.
  105. Shofuda, K. et al. (1997) J. Biol. Chem. 272:9749.
  106. Takino, T. et al. (1995) J. Biol. Chem. 270:23013. NB: This MT-MMP has been re-classified as MT-MMP-3 due to the prior publication of an MT-MMP-2 sequence by Will, H. and Hinzmann, B. (1995) Eur. J. Biochem. 231:602.
  107. Cossins, J. et al. (1996) Biochem. Biophys. Res. Commun. 228:494.
  108. Kolb, C. et al. (1997) Immunol. Lett. 57:83. This is MMP-19 (MMP-18 of Cossins et al.).
  109. Llano, D. et al. (1997) Biochem. 36:15101.