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pH dependence of the enzymatic processing of collagen I by MMP-1 (fibroblast collagenase), MMP-2 (gelatinase A), and MMP-14 ectodomain

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Abstract

The proteolytic processing of collagen I by three matrix metalloproteinases (MMPs), a collagenase (MMP-1), a gelatinase (MMP-2), and the ectodomain of a membrane-type metalloproteinase (MMP-14), has been investigated at 37 °C between pH 6.0 and 9.2, a pH range reflecting conditions found in different body compartments under various physiopathological processes. In the proteolytic degradation the native collagen triple helix must be partially unwound to allow the binding of α chains to the protease’s active-site cleft. We have found that MMP-1 interacts with the two types of collagen I α chains in a similar fashion, whereas both MMP-2 and MMP-14 bind the two α chains in a different way. The overall enzymatic activity is higher on the α-2 chain for both MMP-1 and MMP-2, whereas the MMP-14 ectodomain preferentially cleaves the α-1 chain. In MMP-2 a marked difference for substrate affinity (higher for the α-1 chain) is overwhelmed by an even more marked propensity to cleave the α-2 chain. As a whole, the three classes of MMPs investigated appear to process collagen I in a significantly different fashion, so various MMPs play different roles in the collagen homeostasis in various compartments (such as bloodstream, synovial fluid, normal and tumoral tissues), where different pH values are observed.

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Abbreviations

CD:

Circular dichroism

ECM:

Extracellular matrix

ect-MMP-14:

Ectodomain of matrix metalloproteinase 14

MCA-1:

(7-Methoxycoumarin-4-yl)acetyl-Pro-cyclohexylalanine-Gly-norvaline-His-Ala-(N-3-(2,4-dinitrophenyl)-l-2,3-diaminopropionyl)-NH2

MCA-MMP14:

(7-Methoxycoumarin-4-yl)acetyl-Pro-Leu-Ala-Cys(p-OMeBz)-Trp-Ala-Arg(N-3-(2,4-dinitrophenyl)-l-2,3-diaminopropionyl)-NH2

MCA-omni:

(7-Methoxycoumarin-4-yl)acetyl–Pro-Leu-Gly-Leu-(N-3-(2,4-dinitrophenyl)-l-2,3-diaminopropionyl)-Ala-Arg-NH2

MMP:

Matrix metalloproteinase

PAGE:

Polyacrylamide gel electrophoresis

SDS:

Sodium dodecyl sulfate

Tris:

Tris(hydroxymethyl)aminomethane

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Acknowledgments

This work was financially supported by the Italian Space Agency (ASI 2005 OSMA to U.T. and M.C.). The authors are grateful to Hideaki Nagase and Tayebeh Pourmotabbed for very fruitful discussions.

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Authors and Affiliations

  1. Department of Experimental Medicine and Biochemical Sciences, University of Roma Tor Vergata, Via Montpellier 1, 00133, Rome, Italy

    Magda Gioia, Giovanni Francesco Fasciglione, Susanna Monaco, Diego Sbardella, Stefano Marini & Massimo Coletta

  2. Interuniversity Consortium for the Research on the Chemistry of Metals in Biological Systems, Via C. Ulpiani 27, 70126, Bari, Italy

    Susanna Monaco, Stefano Marini & Massimo Coletta

  3. Department of Surgery, University of Roma Tor Vergata, Via Montpellier 1, 00133, Rome, Italy

    Riccardo Iundusi & Umberto Tarantino

Authors
  1. Magda Gioia
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  2. Giovanni Francesco Fasciglione
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  3. Susanna Monaco
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  4. Riccardo Iundusi
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  5. Diego Sbardella
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  6. Stefano Marini
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  7. Umberto Tarantino
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  8. Massimo Coletta
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Corresponding author

Correspondence to Massimo Coletta.

Additional information

M. Gioia and G. F. Fasciglione contributed equally to the work.

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Gioia, M., Fasciglione, G.F., Monaco, S. et al. pH dependence of the enzymatic processing of collagen I by MMP-1 (fibroblast collagenase), MMP-2 (gelatinase A), and MMP-14 ectodomain. J Biol Inorg Chem 15, 1219–1232 (2010). https://doi.org/10.1007/s00775-010-0680-8

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  • DOI: https://doi.org/10.1007/s00775-010-0680-8

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