title: A caged substrate peptide for matrix metalloproteinases.
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.
authors.affiliation: Max Planck Institute for Chemical Energy Conversion, Stiftstrasse 34-36, D-45470 Mülheim an der Ruhr, Germany. elena.decaneto@cec.mpg.de.
authors.forename: Elena
authors.initials: E
authors.lastname: Decaneto
authors.forename: Stefania
authors.initials: S
authors.lastname: Abbruzzetti
authors.forename: Inge
authors.initials: I
authors.lastname: Heise
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.forename: Cristiano
authors.initials: C
authors.lastname: Viappiani
authors.forename: Markus
authors.initials: M
authors.lastname: Knipp
mesh_terms: D004926:Escherichia coli; D006801:Humans; D006868:Hydrolysis; D013058:Mass Spectrometry; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry* / Q000235:genetics; D020782:Matrix Metalloproteinases / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D055668:Photochemical Processes; D014169:Transformation, Bacterial; D014466:Ultraviolet Rays
chemical_list: D010455:Peptides; D020782:Matrix Metalloproteinases; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14