A simple method for selection of trypsin chromogenic substrates using combinatorial chemistry approach.
A tetrapeptide combinatorial library, considered as chromogenic substrates of bovine beta-trypsin, was synthesized by the solid phase method. The peptides contain an analog of p-nitroanilide, obtained by attaching 5-amino-2-nitrobenzoic acid (Anb(5,2)) to the C-termini. Deconvolution of the peptide library, performed in solution using an iterative method, yielded four efficient trypsin substrates. The most active one, Phe-Val-Pro-Arg-Anb(5,2)-NH(2), appeared to be 125-fold more active than Bz-D,L-Arg-pNA (BAPNA) used as a reference compound. The reported method of designing trypsin chromogenic substrate libraries is straightforward. Such p-nitroanilides may be useful for the investigation of any protease substrate specificity.
Zabłotna
Ewa
E
Faculty of Chemistry, University of Gdańsk, Sobieskiego 18, PL-80-952 Gdańsk, Poland.
Dysasz
Hanna
H
Lesner
Adam
A
Jaśkiewicz
Anna
A
Kaźmierczak
Katarzyna
K
Miecznikowska
Hanna
H
Rolka
Krzysztof
K
D000595:Amino Acid Sequence; D000813:Anilides / Q000737:chemistry; D001671:Biochemistry / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D007700:Kinetics; D008969:Molecular Sequence Data; D009579:Nitrobenzoates / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D014357:Trypsin / Q000737:chemistry*
D000813:Anilides; D009579:Nitrobenzoates; D019151:Peptide Library; D010455:Peptides; C019251:5-amino-2-nitrobenzoic acid; D014357:Trypsin