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Access through your institution Buy or subscribe Matrix metalloproteinases (MMPs) are zinc endopeptidases that are required for the degradation of extracellular matrix components during

normal embryo development, morphogenesis and tissue remodelling1. Their proteolytic activities are precisely regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs)1,2,3,4,5.

Disruption of this balance results in diseases such as arthritis, atherosclerosis, tumour growth and metastasis1,2. Here we report the crystal structure of an MMP-TIMP complex formed

between the catalytic domain of human stromelysin-1 (MMP-3) and human TIMP-1. TIMP-1, a 184-residue protein5, has the shape of an elongated, contiguous wedge. With its long edge, consisting

of five different chain regions, it occupies the entire length of theactive-site cleft of MMP-3. The central disulphide-linked segments Cys 1-Thr 2-Cys 3-Val 4 and Ser 68-Val 69 bind to

either side of the catalytic zinc. Cys 1 bidentally coordinates this zinc, and the Thr-2 side chain extends into the large specificity pocket of MMP-3. This unusual architecture of the

interface between MMP-3 and TIMP-1 suggests new possibilities for designing TIMP variants and synthetic MMP inhibitors with potential therapeutic applications. The TIMP family at present

comprises four members (TIMP-1 to TIMP-4) of relative molecular mass (_M_r) ranging from 22K to 30K, with 40–50% sequence identity3. All TIMPs inhibit active MMPswith relatively low

selectivity, forming tight non-covalent 1:1 complexes4. Native TIMP-1 contains two glycosylation sites5. AC-terminally truncated form of TIMP-1 comprising only the N-terminal two-thirds of

its polypeptide chain (N-TIMP-1) has been shown to inhibit MMP-3 with a slightly reduced affinity compared with full-length TIMP-1 (refs 4,6,7). These experiments suggested that TIMPs

interact with MMPs predominantly through their N-terminal moiety. A preliminary nuclear magnetic resonance (NMR) model of N-TIMP-2 has been reported8, but the mechanism of MMP inhibition by

the TIMPs is not understood. Most MMPs, including stromelysin-1, are secreted as multimodular proenzymes that can be activated, with a 165-residue catalytic domain flanked by an N-terminal

activation peptide and a spacer-linked C-terminal domain, respectively. Three-dimensional structures have been determined for a few catalytic MMP domains, including those of stromelysin-1

(MMP-3)9,10,11 and its proform10, and for full-length porcine MMP-1 (ref. 12). To understand the mechanism of interaction between MMPs and TIMPs, and to allow the design of morespecific TIMP

species, we have undertaken an X-ray crystal-structure analysis of the complex formed between unglycosylated human TIMP-1 and the catalytic domain of MMP-3. This is a preview of

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representation and analysis of surface properties. _Biophys. J._ 64, 166 (1993). Google Scholar  Download references ACKNOWLEDGEMENTS We thank M. Braun for help with crystallization, M. T.

Stubbs for reading the manuscript, and A. Lebedev and E. H. Panepucci for help with molecular replacement. This work was supported by the SFB469, the Human Capital and Mobility, and the

Biotechnology programs of the European Union, the Fonds der Chemischen Industrie, the BMBF, and the NIH. AUTHOR INFORMATION Author notes * Franz-Xaver Gomis-R¨th and Klaus Maskos: These

authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Abteilung für Strukturforschung, Max-Planck-Institut für Biochemie, Martinsried, D-82152, Germany Franz-Xaver Gomis-R¨th,

 Klaus Maskos, Michael Betz, Andreas Bergner, Robert Huber & Wolfram Bode * Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, 66160,

Kansas, USA Ko Suzuki, Naoki Yoshida & Hideaki Nagase * Department of Biochemistry and Molecular Biology, University of Miami, School of Medicine, Miami, 33101, Florida, USA Keith Brew *

AG Proteindynamik MPG-ASMB, c/o DESY, Hamburg, D-22603, Germany Gleb P. Bourenkov & Hans Bartunik Authors * Franz-Xaver Gomis-R¨th View author publications You can also search for this

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can also search for this author inPubMed Google Scholar * Ko Suzuki View author publications You can also search for this author inPubMed Google Scholar * Naoki Yoshida View author

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Google Scholar CORRESPONDING AUTHOR Correspondence to Wolfram Bode. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Gomis-R¨th, FX., Maskos, K., Betz, M.

_et al._ Mechanism of inhibition of the human matrix metalloproteinase stromelysin-1 by TIMP-1. _Nature_ 389, 77–81 (1997). https://doi.org/10.1038/37995 Download citation * Received: 30

April 1997 * Accepted: 13 June 1997 * Issue Date: 04 September 1997 * DOI: https://doi.org/10.1038/37995 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this

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