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A caged substrate peptide for matrix metalloproteinases

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Abstract

Based on the widely applied fluorogenic peptide FS-6 (Mca-Lys-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; Mca = methoxycoumarin-4-acetyl; Dpa = N-3-(2,4-dinitrophenyl)l-α,β-diaminopropionyl) a caged substrate peptide Ac-Lys-Pro-Leu-Gly-Lys*-Lys-Ala-Arg-NH2 (*, position of the cage group) for matrix metalloproteinases was synthesized and characterized. The synthesis implies the modification of a carbamidated lysine side-chain amine with a photocleavable 2-nitrobenzyl group. Mass spectrometry upon UV irradiation demonstrated the complete photolytic cleavage of the protecting group. Time-resolved laser-flash photolysis at 355 nm in combination with transient absorption spectroscopy determined the biphasic decomposition with τa = 171 ± 3 ms (79%) and τb = 2.9 ± 0.2 ms (21%) at pH 6.0 of the photo induced release of the 2-nitrobenzyl group. The recombinantly expressed catalytic domain of human membrane type I matrix metalloproteinase (MT1-MMP or MMP-14) was used to determine the hydrolysis efficiency of the caged peptide before and after photolysis. It turned out that the cage group sufficiently shields the peptide from peptidase activity, which can be thus controlled by UV light.

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Authors and Affiliations

  1. Max Planck Institute for Chemical Energy Conversion, Stiftstrasse 34-36, D-45470, Mülheim an der Ruhr, Germany

    Elena Decaneto, Inge Heise, Wolfgang Lubitz & Markus Knipp

  2. Department of Physical Chemistry II, Ruhr University Bochum, Universitätstrasse 150, D-44780, Bochum, Germany

    Elena Decaneto & Markus Knipp

  3. Department of Life Sciences, University of Parma, Parco Area delle Scienze 11A, I-43124, Parma, Italy

    Stefania Abbruzzetti

  4. Department of Physics and Earth Sciences, University of Parma, Parco Area delle Scienze 7A, I-43124, Parma, Italy

    Cristiano Viappiani

  5. NEST, Istituto Nanoscienze, CNR, Piazza San Silvestro 12, 56127, Pisa, Italy

    Stefania Abbruzzetti & Cristiano Viappiani

Authors
  1. Elena Decaneto
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  2. Stefania Abbruzzetti
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  3. Inge Heise
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  4. Wolfgang Lubitz
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  5. Cristiano Viappiani
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  6. Markus Knipp
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Correspondence to Elena Decaneto or Cristiano Viappiani.

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Decaneto, E., Abbruzzetti, S., Heise, I. et al. A caged substrate peptide for matrix metalloproteinases. Photochem Photobiol Sci 14, 300–307 (2015). https://doi.org/10.1039/c4pp00297k

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