Next Generation of Fluorometric Protease Assays: 7-Nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-Amides) as Class-Spanning Protease Substrates
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Hannah Maus
- Patrick Mueller
- Mergim Meta
- Sabrina N Hoba
- Stefan J Hammerschmidt
- Robert A Zimmermann
- Collin Zimmer
- Natalie Fuchs
- Tanja Schirmeister
- Fabian Barthels
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:001037855700001&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1002/chem.202301855
- eISSN
- 1521-3765
- Externe Identifier
- Clarivate Analytics Document Solution ID: R0RR7
- PubMed Identifier: 37313627
- ISSN
- 0947-6539
- Ausgabe der Veröffentlichung
- 50
- Zeitschrift
- CHEMISTRY-A EUROPEAN JOURNAL
- Schlüsselwörter
- assay interferences
- fluorescent probes
- medicinal chemistry
- nitrobenzofurazane
- proteases
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Titel
- Next Generation of Fluorometric Protease Assays: 7-Nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-Amides) as Class-Spanning Protease Substrates
- Sub types
- Article
- Ausgabe der Zeitschrift
- 29
Datenquelle: Web of Science (Lite)
- Andere Metadatenquellen:
-
- Abstract
- <jats:title>Abstract</jats:title><jats:p>Fluorometric assays are one of the most frequently used methods in medicinal chemistry. Over the last 50 years, the reporter molecules for the detection of protease activity have evolved from first‐generation colorimetric <jats:italic>p</jats:italic>‐nitroanilides, through FRET substrates, and 7‐amino‐4‐methyl coumarin (AMC)‐based substrates. The aim of further substrate development is to increase sensitivity and reduce vulnerability to assay interferences. Herein, we describe a new generation of substrates for protease assays based on 7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl‐amides (NBD‐amides). In this study, we synthesized and tested substrates for 10 different proteases from the serine‐, cysteine‐, and metalloprotease classes. Enzyme‐ and substrate‐specific parameters as well as the inhibitory activity of literature‐known inhibitors confirmed their suitability for application in fluorometric assays. Hence, we were able to present NBD‐based alternatives for common protease substrates. In conclusion, these NBD substrates are not only less susceptible to common assay interference, but they are also able to replace FRET‐based substrates with the requirement of a prime site amino acid residue.</jats:p>
- Autoren
- Hannah Maus
- Patrick Müller
- Mergim Meta
- Sabrina N Hoba
- Stefan J Hammerschmidt
- Robert A Zimmermann
- Collin Zimmer
- Natalie Fuchs
- Tanja Schirmeister
- Fabian Barthels
- DOI
- 10.1002/chem.202301855
- eISSN
- 1521-3765
- ISSN
- 0947-6539
- Ausgabe der Veröffentlichung
- 50
- Zeitschrift
- Chemistry – A European Journal
- Sprache
- en
- Online publication date
- 2023
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Herausgeber
- Wiley
- Herausgeber URL
- http://dx.doi.org/10.1002/chem.202301855
- Datum der Datenerfassung
- 2023
- Titel
- Next Generation of Fluorometric Protease Assays: 7‐Nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl‐amides (NBD‐Amides) as Class‐Spanning Protease Substrates
- Ausgabe der Zeitschrift
- 29
Datenquelle: Crossref
- Abstract
- Fluorometric assays are one of the most frequently used methods in medicinal chemistry. Over the last 50 years, the reporter molecules for the detection of protease activity have evolved from first-generation colorimetric p-nitroanilides, through FRET substrates, and 7-amino-4-methyl coumarin (AMC)-based substrates. The aim of further substrate development is to increase sensitivity and reduce vulnerability to assay interferences. Herein, we describe a new generation of substrates for protease assays based on 7-nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-amides). In this study, we synthesized and tested substrates for 10 different proteases from the serine-, cysteine-, and metalloprotease classes. Enzyme- and substrate-specific parameters as well as the inhibitory activity of literature-known inhibitors confirmed their suitability for application in fluorometric assays. Hence, we were able to present NBD-based alternatives for common protease substrates. In conclusion, these NBD substrates are not only less susceptible to common assay interference, but they are also able to replace FRET-based substrates with the requirement of a prime site amino acid residue.
- Addresses
- Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg-University, Staudingerweg 5, 55128, Mainz, Germany.
- Autoren
- Hannah Maus
- Patrick Müller
- Mergim Meta
- Sabrina N Hoba
- Stefan J Hammerschmidt
- Robert A Zimmermann
- Collin Zimmer
- Natalie Fuchs
- Tanja Schirmeister
- Fabian Barthels
- DOI
- 10.1002/chem.202301855
- eISSN
- 1521-3765
- Externe Identifier
- PubMed Identifier: 37313627
- Funding acknowledgements
- Deutsche Forschungsgemeinschaft: Collaborative Research Center (CRC) SFB 1066 (projects Q5)
- Open access
- false
- ISSN
- 0947-6539
- Ausgabe der Veröffentlichung
- 50
- Zeitschrift
- Chemistry (Weinheim an der Bergstrasse, Germany)
- Schlüsselwörter
- Amides
- Peptide Hydrolases
- Endopeptidases
- Fluorescent Dyes
- Fluorometry
- Sprache
- eng
- Medium
- Print-Electronic
- Online publication date
- 2023
- Paginierung
- e202301855
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Publisher licence
- CC BY
- Datum der Datenerfassung
- 2023
- Titel
- Next Generation of Fluorometric Protease Assays: 7-Nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-Amides) as Class-Spanning Protease Substrates.
- Sub types
- Journal Article
- Ausgabe der Zeitschrift
- 29
Datenquelle: Europe PubMed Central
- Abstract
- Fluorometric assays are one of the most frequently used methods in medicinal chemistry. Over the last 50 years, the reporter molecules for the detection of protease activity have evolved from first-generation colorimetric p-nitroanilides, through FRET substrates, and 7-amino-4-methyl coumarin (AMC)-based substrates. The aim of further substrate development is to increase sensitivity and reduce vulnerability to assay interferences. Herein, we describe a new generation of substrates for protease assays based on 7-nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-amides). In this study, we synthesized and tested substrates for 10 different proteases from the serine-, cysteine-, and metalloprotease classes. Enzyme- and substrate-specific parameters as well as the inhibitory activity of literature-known inhibitors confirmed their suitability for application in fluorometric assays. Hence, we were able to present NBD-based alternatives for common protease substrates. In conclusion, these NBD substrates are not only less susceptible to common assay interference, but they are also able to replace FRET-based substrates with the requirement of a prime site amino acid residue.
- Autoren
- Hannah Maus
- Patrick Müller
- Mergim Meta
- Sabrina N Hoba
- Stefan J Hammerschmidt
- Robert A Zimmermann
- Collin Zimmer
- Natalie Fuchs
- Tanja Schirmeister
- Fabian Barthels
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/37313627
- DOI
- 10.1002/chem.202301855
- eISSN
- 1521-3765
- Funding acknowledgements
- Deutsche Forschungsgemeinschaft: Collaborative Research Center (CRC) SFB 1066 (projects Q5)
- Ausgabe der Veröffentlichung
- 50
- Zeitschrift
- Chemistry
- Schlüsselwörter
- assay interferences
- fluorescent probes
- medicinal chemistry
- nitrobenzofurazane
- proteases
- Peptide Hydrolases
- Amides
- Fluorescent Dyes
- Fluorometry
- Endopeptidases
- Sprache
- eng
- Country
- Germany
- Paginierung
- e202301855
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2023
- Titel
- Next Generation of Fluorometric Protease Assays: 7-Nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-Amides) as Class-Spanning Protease Substrates.
- Sub types
- Journal Article
- Ausgabe der Zeitschrift
- 29
Datenquelle: PubMed
- Author's licence
- CC-BY
- Autoren
- Hannah Maus
- Patrick Müller
- Mergim Meta
- Sabrina N Hoba
- Stefan J Hammerschmidt
- Robert A Zimmermann
- Collin Zimmer
- Natalie Fuchs
- Tanja Schirmeister
- Fabian Barthels
- Hosting institution
- Universitätsbibliothek Mainz
- Sammlungen
- DFG-491381577-H
- Resource version
- Published version
- DOI
- 10.1002/chem.202301855
- Funding acknowledgements
- Deutsche Forschungsgemeinschaft (DFG)|491381577|Open-Access-Publikationskosten 2022–2024 Universität Mainz - Universitätsmedizin
- File(s) embargoed
- false
- Open access
- true
- ISSN
- 1521-3765
- Ausgabe der Veröffentlichung
- 50
- Zeitschrift
- Chemistry - a European journal
- Schlüsselwörter
- 540 Chemie
- 540 Chemistry and allied sciences
- Sprache
- eng
- Open access status
- Open Access
- Paginierung
- e202301855
- Datum der Veröffentlichung
- 2023
- Public URL
- https://openscience.ub.uni-mainz.de/handle/20.500.12030/9532
- Herausgeber
- Wiley-VCH
- Datum der Datenerfassung
- 2023
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2023
- Zugang
- Public
- Titel
- Next generation of fluorometric protease assays : 7-nitrobenz-2-oxa-1,3-diazol-4-yl-amides (NBD-amides) as class-spanning protease substrates
- Ausgabe der Zeitschrift
- 29
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