Hydrosoluble fluorogenic substrates for plasmin.
New hydrosoluble fluorogenic substrates for plasmin gluconoylpeptidyl-3-amido-9-ethylcarbazole were synthesized. The substitution of the N-terminal end of the peptides by a gluconoyl group prevents the substrates from aminopeptidase degradation and highly increases their hydrosolubility. The substitution of the peptide C-terminal end by a 3-amino-9-ethylcarbazole group leads to substrates suitable for direct fluorometric assay of plasmin present in cell supernatants or in cell lysates. On the basis of the kinetic parameters of the substrate hydrolysis by plasmin, it was found that D amino acids in the P2 position decrease systematically the kinetic constants of the substrates. The L configuration of the P2 amino acid appears therefore as essential in optimum substrates for plasmin.
Harnois-Pontoni
M
M
Département de Biochimie des Glycoconjugés et Lectines Endogènes, Centre de Biophysique Moléculaire, CNRS, Orléans, France.
Monsigny
M
M
Mayer
R
R
D000595:Amino Acid Sequence; D000818:Animals; D002227:Carbazoles / Q000138:chemical synthesis*; D002851:Chromatography, High Pressure Liquid; D005341:Fibrinolysin / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D006801:Humans; D007536:Isomerism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis*; D010446:Peptide Fragments / Q000737:chemistry; D012995:Solubility; D013379:Substrate Specificity
D002227:Carbazoles; D005456:Fluorescent Dyes; D009842:Oligopeptides; D010446:Peptide Fragments; C020702:3-amino-9-ethylcarbazole; D005341:Fibrinolysin