Two Tags in One Probe: Combining Fluorescence- and Biotin-based Detection of the Trypanosomal Cysteine Protease Rhodesain

Publikationstyp:
Zeitschriftenaufsatz
Metadaten:
Autoren
  • Carina Lemke
  • Adela Jilkova
  • Dominic Ferber
  • Annett Braune
  • Anja On
  • Patrick Johe
  • Alena Zikova
  • Tanja Schirmeister
  • Michael Mares
  • Martin Horn
  • Michael Gutschow
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000850603500001&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1002/chem.202201636
eISSN
1521-3765
Externe Identifier
  • Clarivate Analytics Document Solution ID: 5Y9ET
  • PubMed Identifier: 35852812
ISSN
0947-6539
Ausgabe der Veröffentlichung
62
Zeitschrift
CHEMISTRY-A EUROPEAN JOURNAL
Schlüsselwörter
  • activity-based probes
  • affinity chromatography
  • cysteine proteases
  • imaging agents
  • peptidomimetics
  • rhodesain
  • size exclusion chromatography
Artikelnummer
ARTN e202201636
Datum der Veröffentlichung
2022
Status
Published
Titel
Two Tags in One Probe: Combining Fluorescence- and Biotin-based Detection of the Trypanosomal Cysteine Protease Rhodesain
Sub types
  • Article
Ausgabe der Zeitschrift
28

Datenquelle: Web of Science (Lite)

Andere Metadatenquellen:
Abstract
<jats:title>Abstract</jats:title><jats:p>Rhodesain is the major cysteine protease of the protozoan parasite <jats:italic>Trypanosoma brucei</jats:italic> and a therapeutic target for sleeping sickness, a fatal neglected tropical disease. We designed, synthesized and characterized a bimodal activity‐based probe that binds to and inactivates rhodesain. This probe exhibited an irreversible mode of action and extraordinary potency for the target protease with a <jats:italic>k</jats:italic><jats:sub>inac</jats:sub>/<jats:italic>K</jats:italic><jats:sub>i</jats:sub> value of 37,000 M<jats:sup>−1</jats:sup>s<jats:sup>−1</jats:sup>. Two reporter tags, a fluorescent coumarin moiety and a biotin affinity label, were incorporated into the probe and enabled highly sensitive detection of rhodesain in a complex proteome by in‐gel fluorescence and on‐blot chemiluminescence. Furthermore, the probe was employed for microseparation and quantification of rhodesain and for inhibitor screening using a competition assay. The developed bimodal rhodesain probe represents a new proteomic tool for studying <jats:italic>Trypanosoma</jats:italic> pathobiochemistry and antitrypanosomal drug discovery.</jats:p>
Autoren
  • Carina Lemke
  • Adéla Jílková
  • Dominic Ferber
  • Annett Braune
  • Anja On
  • Patrick Johe
  • Alena Zíková
  • Tanja Schirmeister
  • Michael Mareš
  • Martin Horn
  • Michael Gütschow
DOI
10.1002/chem.202201636
eISSN
1521-3765
ISSN
0947-6539
Ausgabe der Veröffentlichung
62
Zeitschrift
Chemistry – A European Journal
Sprache
en
Online publication date
2022
Datum der Veröffentlichung
2022
Status
Published
Herausgeber
Wiley
Herausgeber URL
http://dx.doi.org/10.1002/chem.202201636
Datum der Datenerfassung
2023
Titel
Two Tags in One Probe: Combining Fluorescence‐ and Biotin‐based Detection of the Trypanosomal Cysteine Protease Rhodesain
Ausgabe der Zeitschrift
28

Datenquelle: Crossref

Abstract
Rhodesain is the major cysteine protease of the protozoan parasite Trypanosoma brucei and a therapeutic target for sleeping sickness, a fatal neglected tropical disease. We designed, synthesized and characterized a bimodal activity-based probe that binds to and inactivates rhodesain. This probe exhibited an irreversible mode of action and extraordinary potency for the target protease with a k<sub>inac</sub> /K<sub>i</sub> value of 37,000 M<sup>-1</sup> s<sup>-1</sup> . Two reporter tags, a fluorescent coumarin moiety and a biotin affinity label, were incorporated into the probe and enabled highly sensitive detection of rhodesain in a complex proteome by in-gel fluorescence and on-blot chemiluminescence. Furthermore, the probe was employed for microseparation and quantification of rhodesain and for inhibitor screening using a competition assay. The developed bimodal rhodesain probe represents a new proteomic tool for studying Trypanosoma pathobiochemistry and antitrypanosomal drug discovery.
Addresses
  • Pharmaceutical Institute, Department of Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
Autoren
  • Carina Lemke
  • Adéla Jílková
  • Dominic Ferber
  • Annett Braune
  • Anja On
  • Patrick Johe
  • Alena Zíková
  • Tanja Schirmeister
  • Michael Mareš
  • Martin Horn
  • Michael Gütschow
DOI
10.1002/chem.202201636
eISSN
1521-3765
Externe Identifier
  • PubMed Identifier: 35852812
  • PubMed Central ID: PMC9826439
Open access
true
ISSN
0947-6539
Ausgabe der Veröffentlichung
62
Zeitschrift
Chemistry (Weinheim an der Bergstrasse, Germany)
Schlüsselwörter
  • Trypanosoma
  • Trypanosoma brucei brucei
  • Biotin
  • Proteomics
  • Structure-Activity Relationship
  • Fluorescence
  • Cysteine Proteases
Sprache
eng
Medium
Print-Electronic
Online publication date
2022
Open access status
Open Access
Paginierung
e202201636
Datum der Veröffentlichung
2022
Status
Published
Publisher licence
CC BY-NC-ND
Datum der Datenerfassung
2022
Titel
Two Tags in One Probe: Combining Fluorescence- and Biotin-based Detection of the Trypanosomal Cysteine Protease Rhodesain.
Sub types
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
28

Files

https://europepmc.org/articles/PMC9826439?pdf=render

Datenquelle: Europe PubMed Central

Abstract
Rhodesain is the major cysteine protease of the protozoan parasite Trypanosoma brucei and a therapeutic target for sleeping sickness, a fatal neglected tropical disease. We designed, synthesized and characterized a bimodal activity-based probe that binds to and inactivates rhodesain. This probe exhibited an irreversible mode of action and extraordinary potency for the target protease with a kinac /Ki value of 37,000 M-1 s-1 . Two reporter tags, a fluorescent coumarin moiety and a biotin affinity label, were incorporated into the probe and enabled highly sensitive detection of rhodesain in a complex proteome by in-gel fluorescence and on-blot chemiluminescence. Furthermore, the probe was employed for microseparation and quantification of rhodesain and for inhibitor screening using a competition assay. The developed bimodal rhodesain probe represents a new proteomic tool for studying Trypanosoma pathobiochemistry and antitrypanosomal drug discovery.
Autoren
  • Carina Lemke
  • Adéla Jílková
  • Dominic Ferber
  • Annett Braune
  • Anja On
  • Patrick Johe
  • Alena Zíková
  • Tanja Schirmeister
  • Michael Mareš
  • Martin Horn
  • Michael Gütschow
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/35852812
DOI
10.1002/chem.202201636
eISSN
1521-3765
Externe Identifier
  • PubMed Central ID: PMC9826439
Ausgabe der Veröffentlichung
62
Zeitschrift
Chemistry
Schlüsselwörter
  • activity-based probes
  • affinity chromatography
  • cysteine proteases
  • imaging agents
  • peptidomimetics
  • rhodesain
  • size exclusion chromatography
  • Biotin
  • Cysteine Proteases
  • Fluorescence
  • Proteomics
  • Trypanosoma brucei brucei
  • Trypanosoma
  • Structure-Activity Relationship
Sprache
eng
Country
Germany
Paginierung
e202201636
Datum der Veröffentlichung
2022
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2022
Titel
Two Tags in One Probe: Combining Fluorescence- and Biotin-based Detection of the Trypanosomal Cysteine Protease Rhodesain.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
28

Datenquelle: PubMed

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