Fluorescence-Based On-Resin Detection of Three Model Proteases
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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-12-01
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| Series: | Chemosensors |
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| Online Access: | https://www.mdpi.com/2227-9040/9/12/359 |
| _version_ | 1797505920570228736 |
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| author | David Milićević Jan Hlaváč |
| author_facet | David Milićević Jan Hlaváč |
| author_sort | David Milićević |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T04:25:13Z |
| format | Article |
| id | doaj.art-d18cf45da527471088370202acf6e443 |
| institution | Directory Open Access Journal |
| issn | 2227-9040 |
| language | English |
| last_indexed | 2024-03-10T04:25:13Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Chemosensors |
| spelling | doaj.art-d18cf45da527471088370202acf6e4432023-11-23T07:41:36ZengMDPI AGChemosensors2227-90402021-12-0191235910.3390/chemosensors9120359Fluorescence-Based On-Resin Detection of Three Model ProteasesDavid Milićević0Jan Hlaváč1Department of Organic Chemistry, Faculty of Science, Palacký University, Tř. 17. Listopadu 12, 771 46 Olomouc, Czech RepublicDepartment of Organic Chemistry, Faculty of Science, Palacký University, Tř. 17. Listopadu 12, 771 46 Olomouc, Czech RepublicA 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.https://www.mdpi.com/2227-9040/9/12/359solid-phase synthesisimmobilized probeon-resin peptide cleavagemodel systemsimultaneous detectionprotease |
| spellingShingle | David Milićević Jan Hlaváč Fluorescence-Based On-Resin Detection of Three Model Proteases Chemosensors solid-phase synthesis immobilized probe on-resin peptide cleavage model system simultaneous detection protease |
| title | Fluorescence-Based On-Resin Detection of Three Model Proteases |
| title_full | Fluorescence-Based On-Resin Detection of Three Model Proteases |
| title_fullStr | Fluorescence-Based On-Resin Detection of Three Model Proteases |
| title_full_unstemmed | Fluorescence-Based On-Resin Detection of Three Model Proteases |
| title_short | Fluorescence-Based On-Resin Detection of Three Model Proteases |
| title_sort | fluorescence based on resin detection of three model proteases |
| topic | solid-phase synthesis immobilized probe on-resin peptide cleavage model system simultaneous detection protease |
| url | https://www.mdpi.com/2227-9040/9/12/359 |
| work_keys_str_mv | AT davidmilicevic fluorescencebasedonresindetectionofthreemodelproteases AT janhlavac fluorescencebasedonresindetectionofthreemodelproteases |