title: Synthesis and Kinetic Characterisation of Water-Soluble Fluorogenic Acyl Donors for Transglutaminase 2.
abstract: Small glutamate-containing peptides bearing coumarin derivatives as fluorescent leaving groups attached to the γ-carboxylic acid group of the Glu residue were synthesised and investigated with regard to their potential to act as substrates for transglutaminase 2 (TGase 2). Their synthesis was accomplished by an efficient solid-phase approach. The excellent water solubility of the compounds enabled their extensive kinetic characterisation in the context of TGase 2-catalysed hydrolysis and aminolysis. The influence of the coumarin skeleton's substitution pattern on the kinetic properties was studied. Derivatives containing 7-hydroxy-4-methylcoumarin (HMC) revealed properties superior to those of their 7-hydroxycoumarin counterparts; analogous amides are not accepted as substrates. Z-Glu(HMC)-Gly-OH, which exhibited the best substrate properties out of the investigated derivatives, was selected for representative kinetic characterisation of acyl acceptor substrates and irreversible inhibitors.
authors.affiliation: Helmholtz-Zentrum Dresden-Rossendorf, Institut für Radiopharmazeutische Krebsforschung, Bautzner Landstrasse 400, 01328, Dresden, Germany.|Fachrichtung Chemie und Lebensmittelchemie, Technische Universität Dresden, Mommsenstrasse 4, 01062, Dresden, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Wodtke
authors.affiliation: Helmholtz-Zentrum Dresden-Rossendorf, Institut für Radiopharmazeutische Krebsforschung, Bautzner Landstrasse 400, 01328, Dresden, Germany.|Department of Imaging and Pathology, KU Leuven, UZ Herestraat 49, 3000, Leuven, Belgium.
authors.forename: Georg
authors.initials: G
authors.lastname: Schramm
authors.affiliation: Helmholtz-Zentrum Dresden-Rossendorf, Institut für Radiopharmazeutische Krebsforschung, Bautzner Landstrasse 400, 01328, Dresden, Germany.|Fachrichtung Chemie und Lebensmittelchemie, Technische Universität Dresden, Mommsenstrasse 4, 01062, Dresden, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Pietzsch
authors.affiliation: Zentrum für Pharmakologie, Medizinische Fakultät, Universität zu Köln, Gleueler Strasse 24, 50931, Köln, Germany. markus.pietsch@uk-koeln.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Pietsch
authors.affiliation: Helmholtz-Zentrum Dresden-Rossendorf, Institut für Radiopharmazeutische Krebsforschung, Bautzner Landstrasse 400, 01328, Dresden, Germany. r.loeser@hzdr.de.|Fachrichtung Chemie und Lebensmittelchemie, Technische Universität Dresden, Mommsenstrasse 4, 01062, Dresden, Germany. r.loeser@hzdr.de.
authors.forename: Reik
authors.initials: R
authors.lastname: Löser
mesh_terms: D000588:Amines / Q000737:chemistry; D000607:Aminoacetonitrile / Q000737:chemistry; D000818:Animals; D000975:Antioxidants / Q000737:chemistry; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D003374:Coumarins / Q000138:chemical synthesis / Q000737:chemistry*; D004229:Dithiothreitol / Q000737:chemistry; D057075:Enzyme Assays / Q000379:methods; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D019204:GTP-Binding Proteins / Q000737:chemistry*; D005971:Glutamates / Q000138:chemical synthesis / Q000737:chemistry; D006168:Guinea Pigs; D006801:Humans; D007460:Iodoacetamide / Q000737:chemistry; D007700:Kinetics; D008239:Lysine / Q000031:analogs & derivatives / Q000737:chemistry; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry*; D010720:Phosphines / Q000737:chemistry; D010879:Piperazines / Q000737:chemistry; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases / Q000737:chemistry*
chemical_list: D000588:Amines; D000607:Aminoacetonitrile; D000975:Antioxidants; D001710:Biotin; D003374:Coumarins; D004229:Dithiothreitol; D005456:Fluorescent Dyes; D019204:GTP-Binding Proteins; D005971:Glutamates; D007460:Iodoacetamide; D008239:Lysine; D009842:Oligopeptides; D010720:Phosphines; D010879:Piperazines; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; C000622620:Nalpha-phenylacetyl-Nepsilon-acryloyllysine-4-(6-methylpyridine-2-yl)piperazide; C076022:5-(biotinamido)pentylamine; C080938:tris(2-carboxyethyl)phosphine
keywords: enzyme assays; inhibitors; reactive peptides; solid-phase synthesis; transferases