Pressure and Temperature Effects on the Activity and Structure of the Catalytic Domain of Human MT1-MMP |
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| Authors: | Elena Decaneto Saba Suladze Christopher Rosin Martina Havenith Wolfgang Lubitz Roland Winter |
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| Institution: | 1Max Planck Institute for Chemical Energy Conversion, Mülheim a. d. Ruhr, Germany;2Department of Chemistry and Chemical Biology, Physical Chemistry, Technische Universität Dortmund, Dortmund, Germany;3Department of Physical Chemistry II, Ruhr-University Bochum, Bochum, Germany |
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| Abstract: | Membrane type 1-matrix metalloproteinase (MT1-MMP or MMP-14) is a zinc-transmembrane metalloprotease involved in the degradation of extracellular matrix and tumor invasion. While changes in solvation of MT1-MMP have been recently studied, little is known about the structural and energetic changes associated with MT1-MMP while interacting with substrates. Steady-state kinetic and thermodynamic data (including activation energies and activation volumes) were measured over a wide range of temperatures and pressures by means of a stopped-flow fluorescence technique. Complementary temperature- and pressure-dependent Fourier-transform infrared measurements provided corresponding structural information of the protein. MT1-MMP is stable and active over a wide range of temperatures (10–55°C). A small conformational change was detected at 37°C, which is responsible for the change in activity observed at the same temperature. Pressure decreases the enzymatic activity until complete inactivation occurs at 2 kbar. The inactivation is associated with changes in the rate-limiting step of the reaction caused by additional hydration of the active site upon compression and/or minor conformational changes in the active site region. Based on these data, an energy and volume diagram could be established for the various steps of the enzymatic reaction. |
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