Synthesis of N epsilon-(7-diethylaminocoumarin-3-carboxyl)- and N epsilon-(7-methoxycoumarin-3-carboxyl)-L-Fmoc lysine as tools for protease cleavage detection by fluorescence.
Two coumarin-labelled lysines were conveniently prepared as a fluorescence resonance energy transfer (FRET) pair for peptide cleavage detection. 7-Methoxy and 7-diethylamino coumarin-3-carboxylic acids were synthesized according to a modification of known procedures. Labelling at lysine was achieved in solution via the active N-hydroxysuccinimide ester of the carboxylic acid coumarin derivatives to give the target compounds in good yield. Subsequently, these modified amino acids were used in solid phase peptide synthesis (SPPS), and their potential utility in an extracellular matrix metalloprotease (MMP-1) activity measurement via FRET and/or quenching studies was demonstrated.
Berthelot
Thomas
T
Groupe de Chimie Bio-Organique, Université Victor Segalen Bordeaux 2, France.
Talbot
Jean-Claude
JC
Laïn
Georges
G
Déleris
Gérard
G
Latxague
Laurent
L
D002851:Chromatography, High Pressure Liquid; D003374:Coumarins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D007700:Kinetics; D008239:Lysine / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D015394:Molecular Structure
D003374:Coumarins; C505106:N(epsilon)-(7-diethylaminocoumarin-3-carboxyl)-9-fluorenylmethoxycarbonyllysine; C505105:N(epsilon)-(7-methoxycoumarin-3-carboxyl)-9-fluorenylmethoxycarbonyllysine; D020781:Matrix Metalloproteinase 1; D008239:Lysine