PMID: 3771103Aug 1, 1986Paper

Fluorogenic substrates for chymotrypsin with new fluorescent markers

International Journal of Peptide and Protein Research
G Kokotos, C Tzougraki

Abstract

Several substrates for chymotrypsin Glt-Phe-NHRx have been synthesized, where RxNH2 are the compounds 3-aminocoumarin (2), 6-aminocoumarin (3), 3-acetamido-6-aminocoumarin (4), 3-acetamido-8-aminocoumarin (5), 7-amino-4-methyl-2-quinolinone (AMQ), (6). The fluorescence properties of the new substrates and those of the corresponding free amines were examined. The compound 7-glutarylphenylalaninamido-4-methyl-2-quinolinone (Glt-Phe-AMQ), (15), provided a new suitable substrate for chymotrypsin determination. The enzymatic release of the fluorophore AMQ was measured at lambda ex = 360 nm and lambda em = 435 nm. The Km of 15 was 0.5 mM and its kcat/Km ratio was 47 M-1 s-1. By using this substrate, the detection limit of chymotrypsin was 10 ng/ml.

References

Nov 15, 1979·Analytical Biochemistry·G D SmithT J Peters
Feb 1, 1977·Chemical & Pharmaceutical Bulletin·Y KanaokaH Nakayama
Jan 1, 1976·Analytical Biochemistry·M ZimmermanG Patel
Jan 1, 1981·Analytical Biochemistry·K MurakamiS Sakakibara
Apr 1, 1982·Chemical & Pharmaceutical Bulletin·Y KanaokaT Sekine

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Citations

Jan 11, 2001·The Journal of Peptide Research : Official Journal of the American Peptide Society·C CharitosG Kokotos
Sep 1, 1990·Biological Chemistry Hoppe-Seyler·G KokotosC Tzougraki
Jan 1, 1997·The Journal of Peptide Research : Official Journal of the American Peptide Society·C NoulaC Tzougraki

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