Fluorescence-Based On-Resin Detection of Three Model Proteases

Chemosensors

Jan Hlaváč¹  David Milićević¹ 

[1] Department of Organic Chemistry, Faculty of Science, Palacký University, Tř. 17. Listopadu 12, 771 46 Olomouc, Czech Republic;

关键词: solid-phase synthesis;    immobilized probe;    on-resin peptide cleavage;    model system;    simultaneous detection;    protease;   

DOI  :  10.3390/chemosensors9120359

来源: DOAJ

【 摘 要 】

A new approach to on-resin detection of three model proteases (trypsin, chymotrypsin, and thrombin) has been developed, while at the same time already described methodology for simultaneous detection of two enzymes (trypsin and chymotrypsin) has been additionally generalized. Appropriate immobilized substrates, comprising specifically cleavable peptide sequences capped with fluorescent dyes, have been synthesized on Rink Amide PEGA resin or Amino PEGA resin modified with backbone amide linker (BAL). Resulting solid support-bound probes were then dispersed into Tris-HCl buffer solution (pH = 8.0) and subjected to enzymatic cleavage. Liberated fluorophores have been tracked by fluorescence measuring. The competitive activities of studied proteases towards the thrombin probe have been efficiently limited and controlled by employing a Bowman-Birk inhibitor into a system.

【 授权许可】

Unknown