Cell Science at a Glance

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extracellular matrix. Besides modulating tissue structure to facilitate remodeling, cell migration, etc. MMP activities can result in the generation of epitopes that act as cell effectors or the release of sequestered growth factors. Proteolysis of adhesion molecules, growth factors, cytokines, chemokines and receptors have all been documented (Sternlicht and Werb, 2001) and the potential effects on cell behaviour are multifarious. There are 24 human MMPs, and homologues can be identied in birds, African clawed toad (Xenopus laevis), zebrash (Danio rerio), fruit y (Drosophila melanogaster), sea urchin (Paracentrotus lividus), nematode (Caenorhabditis elegans) and Hydra (Hydra vulgaris), as well as in plants and algae. Because of their recognized role in disease the MMPs have long been considered as pharmacological targets, but their multiplicity, associated with their variable expression in different tissues and their apparently overlapping substrate specicities, has presented considerable challenges to those hoping to design suitable therapeutic inhibitors. As a consequence of their apparent redundancy, the majority of studies in which MMP genes have been ablated in mice have produced no overt or very subtle phenotypes, with the exception of MMP-14, which, when knocked out, gave defects in endochondral and intramembranous bone development. However, specic challenges to individual knockouts are yielding a clearer picture of novel tissue functions of the MMPs, at the level of both cell-cell and cell-matrix interactions (Sternlicht and Werb, 2001). The MMPs are zinc-dependent

Matrix metalloproteinases at a glance

Meng-Huee Lee and Gillian Murphy*
Dept of Oncology, University of Cambridge, Cambridge Institute for Medical Research, Hills Road, Cambridge CB2 2XY, UK *Author for correspondence (e-mail: gm290@cam.ac.uk)
Journal of Cell Science 117, 4015-4016 Published by The Company of Biologists 2004 doi:10.1242/jcs.01223

The matrix metalloproteinases (MMPs) are one of the major families of proteinases that play key roles in the responses of cells to their microenvironment. Most notably the MMPs have the combined capacity to degrade all the components of the

Meng-Huee Lee and Gillian Murphy

Catalytic domain
Pro

Fibronectin-like repeats

Hemopexin-like domain

Soluble MMPs
Zn

Matrilysins MMP-7, MMP-26

CAT Pro
Zn

HP

Collagenases (MMP-1, MMP-8, MMP-13) Stromelysins (MMP-3, MMP-10) Metalloelastase (MMP-12) MMP-19, enamelysin (MMP-20) MMP-27 Stromelysin 3 (MMP-11) MMP-21 Epilysin (MMP-28)

CAT Pro
Zn

S HP

CAT Pro

S HP

S
jcs.biologists.org

MMP2

MMP2

MMP2

Gelatinases (MMP-2 and MMP-9)

Zn

CAT

Regulation, localisation and functions of MMPs

TIMP-1, TIMP-2, TIMP-3 and TIMP-4

Membrane-associated MMPs
Membrane-type (MT-1) MMP, MT-2 MMP, MT-3 MMP, MT-5 MMP, MMP-5 (MMP-14, MMP-15, MMP-16 and MMP-24)

Pro
Zn

HP TM CD

Cell-cell interactions
Proteinase activation cascades S Proteinase inhibitor processing e.g. 1proteinase inhibitor Release of membrane growth factors e.g. TGF- S GPI Processing of chemokines Shedding of growth factor receptors e.g. TNFR-I and TNFR-II S CA Ig Shedding of adhesion molecules e.g. L-selectin, cadherins and CD44 ECM-bound or EC-milieu MMPs Endocytosis? Secretion/ activation Cell surface MMPs TIMP inhibition MMP

CAT Pro

S HP

MT-4 MMP, MT-6 MMP (MMP-17 and MMP-25)

Zn

CAT Pro TM

S HP

CA MMP (MMP-23)

Zn

CAT

Golgi apparatus

Cell-matrix interactions
Pro HP

Propeptide
CA

Zn

ECM dissolution e.g. tumour metastasis

S

CAT

Catalytic domain Fibronectin-like repeats

Hemopexin-like domain

Cysteine array

TM Transmembrane domain
CD

Release of bioactive fragments Release of growth factors

Gene transcription

Prohormone convertase cleavage domain

GPI

Cytoplasmic tail Hinge

Nucleus

GPI anchor

Ig

Immunoglobulin C2-type fold

Journal of Cell Science 2004 (117, pp. 4015-4016)

(See poster insert)

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Journal of Cell Science 117 (18)

specic biological relevance. The other MMP domains have distinct functions, such as as exosites for substrate interactions, e.g. the hemopexin-like domains of MMP-1, MMP-8, MMP-13, MMP-14, MMP-16 and MMP-18 are essential for their ability to cleave brillar collagens and the bronectin-like domains of MMP-2 and MMP-9 confer their binding to denatured collagen substrates. The hemopexin-like domain of MMP-14 can homodimerise in order to promote its clustering at the cell surface, a property that promotes its activity. The hemopexin-like domain confers the ability to interact with other extracellular matrix components and cell adhesion molecules and may be of signicance in the determination of specic pericellular locations of individual MMPs. The MMPs are regulated at the transcriptional and post-transcriptional levels, as well as by activation, inhibition and cell/ECM localization, which allows tissue-specic spatial and temporal patterns of functional activity. Expression levels may be modulated by different cytokines, growth factors, hormones, extracellular matrix interactions and cytoskeletal changes through specic elements in the MMP promoters governing transcriptional regulation. Sequestration of the secreted MMPs in Golgi vesicles has been described for many stimulated cells, as has storage of MMP-8 and MMP-9 in the secretory granules of PMN leucocytes. The membrane-associated MMPs appear to have distinct trafficking pathways to specic sites at the cell surface. Association of some MMPs with integrins and other cell surface receptors has been described, e.g. MMP1integrin-21, MMP-2integrinV3, MMP-14integrin-21/V3, MMP-7CD44 and MMP-9CD44. Many MMPs bind to specic ECM components (see above). With the exception of very rapidly remodeling tissues, extracellular levels of MMPs tend to be quite low, and unambiguous immunohistochemical detection is challenging. The four TIMPs act as a further level of extracellular regulation and also have specic patterns of gene regulation and tissue-specic expression. TIMP-3 is unusual in that it is largely sequestered into the extracellular matrix or at the cell surface via heparan sulphate proteoglycans. Individual TIMPs differ in their ability to inhibit different MMPs; TIMP-1 is a poor inhibitor of MMP-14, MMP-16 and MMP-19. In addition there are specic interactions of TIMP-1 with proMMP-9, of TIMP-2 with proMMP-2 and of TIMP-3 with both proMMP-2 and proMMP-9 by binding through their three C-terminal disulphide-bonded loops, which allows complexes of the inactive MMPs to be formed, as well as giving very tight-binding active enzyme complexes. The true signicance of this has only been elucidated for proMMP-2, where the TIMP-2 complex allows binding of the MMP to MMP-14 at the cell surface, promoting its activation and potentially focusing proteolysis to specic sites. The activation of proMMPs in general is probably strictly pericellular, e.g. where plasmin, generated by the activity of urokinase-type plasminogen activator, is an initiator of activation cascades. If there is an excess of TIMPs and serine proteinase inhibitors in the environment, these may also conne activity to the local environment. There is a further level of regulation of the MMPs through clearance by endocytosis. Little is known of the fate of most MMP-TIMP complexes, but complexes with 2 macroglobulin are thought to be endocytosed after binding to the low density lipoprotein receptor related protein (LRP). Thrombospondin 2 modulates both MMP-9TIMP-1 and MMP-2 internalisation via LRP. The membrane-associated proteinase MMP14 is endocytosed via clathrin- and nonclathrin-mediated pathways and may recycle to the cell surface in some situations. The other MT-MMPs probably have similar properties.

endopeptidases of the superfamily Metzincins (MEROPS, the protease database). They have specic domain structures, minimally consisting of a propeptide and a catalytic domain (MMP-7 and MMP-26), commonly with the addition of a hemopexin-like, fourbladed propeller domain connected by a linker or hinge region (MMP-1, MMP3, MMP-8, MMP-11, MMP-12, MMP13, MMP-18, MMP-19, MMP-20, MMP-21, MMP-27 and MMP-28). Others have these features plus a bronectin-like domain of three type II repeats (MMP-2 and MMP-9) or a transmembrane region and a short cytoplasmic tail (MMP-14, MMP-15, MMP-16 and MMP-24), or a glycosylphosphatidyl anchor (MMP-17 and MMP-25). MMP-23 is exceptional in that it has unique cysteine-rich, proline-rich and IL-1 receptor type II like domains and might initially be anchored by an N-terminal transmembrane domain prior to propeptide processing. The propeptide of the MMPs contains a cysteine switch motif, PRCGXPD, in which the cysteine residue interacts with the catalytic zinc domain in order to maintain inactivity until the propeptide has been removed by proteolysis. The catalytic domains have the zinc-binding motif HEXGHXXGXXH, in which the three histidine residues ligate the zinc ion. Activation of the MMPs by propeptide removal is a critical feature of their regulation and, in the case of MMPs with the requisite RX(R/K)R motif (MMP-11, MMP-14, MMP-15, MMP16, MMP-21, MMP-23, MMP-24, MMP-25 and MMP-28), might be effected intracellularly by the action of trans-Golgi-localised proprotein convertases or, for the majority, by cleavage by plasmin, autolysis or the action of other MMPs at the cell surface. MMP activity may subsequently be regulated by the action of inhibitors, notably the tissue inhibitors of MMPs (TIMPs) TIMP-1, TIMP-2, TIMP-3 and TIMP-4 and the serum panproteinase inhibitor 2 macroglobulin (Baker et al., 2002) The TIMPs are sixloop disulphide-bonded proteins forming two domains. They interact via their Nterminal three disulphide-bonded loops with the active site cleft of the catalytic domain, although signicant interactions of the hemopexin-like domains of MMP2 and MMP-9 with the C-terminal domains of TIMPs appear to have

Recommended reading
Sternlicht, M. D. and Werb, Z. (2001). How matrix metalloproteinases regulate cell behavior. Annu. Rev. Cell Dev. Biol. 17, 463-516. Baker, A. H., Edwards, D. R. and Murphy, G. (2002). Metalloproteinase inhibitors: biological actions and therapeutic opportunities. J. Cell Sci. 115, 3719-3727. MEROPS: the Protease Database [http://merops. sanger.ac.uk].

Cell Science at a Glance on the Web Electronic copies of the poster insert are available in the online version of this article at jcs.biologists.org. The JPEG images can be downloaded for printing or used as slides.