title: Pressure and Temperature Effects on the Activity and Structure of the Catalytic Domain of Human MT1-MMP.
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.
authors.affiliation: Max Planck Institute for Chemical Energy Conversion, Mülheim a. d. Ruhr, Germany; Department of Physical Chemistry II, Ruhr-University Bochum, Bochum, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Decaneto
authors.affiliation: Department of Chemistry and Chemical Biology, Physical Chemistry, Technische Universität Dortmund, Dortmund, Germany.
authors.forename: Saba
authors.initials: S
authors.lastname: Suladze
authors.affiliation: Department of Chemistry and Chemical Biology, Physical Chemistry, Technische Universität Dortmund, Dortmund, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Rosin
authors.affiliation: Department of Physical Chemistry II, Ruhr-University Bochum, Bochum, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Havenith
authors.affiliation: Max Planck Institute for Chemical Energy Conversion, Mülheim a. d. Ruhr, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.affiliation: Department of Chemistry and Chemical Biology, Physical Chemistry, Technische Universität Dortmund, Dortmund, Germany. Electronic address: roland.winter@tu-dortmund.de.
authors.forename: Roland
authors.initials: R
authors.lastname: Winter
mesh_terms: D020134:Catalytic Domain; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry* / Q000378:metabolism*; D011312:Pressure; D017433:Protein Structure, Secondary; D013696:Temperature; D013816:Thermodynamics
chemical_list: C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14