Publications Repository - Helmholtz-Zentrum Dresden-Rossendorf

Synthesis and kinetic characterisation of water-soluble fluorogenic acyl donors for transglutaminase 2

Wodtke, R.; Schramm, G.; Pietzsch, J.; Pietsch, M.; Löser, R.

Small glutamate-containing peptides that bear coumarin derivatives as fluorescent leaving groups attached to the gamma-carboxylic group of the Glu residue were synthesised and investigated towards 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 regarding TGase 2-catalysed hydrolysis and aminolysis. The influence of the substitution pattern at the coumarin skeleton on the kinetic properties was studied. Derivatives containing 7-hydroxy-4-methylcoumarin (HMC) revealed superior properties over their 7-hydroxycoumarin counterparts; analogous amides are not accepted as substrates. Z-Glu(HMC)-Gly-OH, which exhibited the most optimal substrate properties among the investigated derivatives, was selected for the exemplary kinetic characterisation of acyl acceptor substrates and irreversible inhibitors.

Keywords: enzyme assays; inhibitors; reactive peptides; solid-phase synthesis; transferases


Publ.-Id: 23553