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

Overview

Journal Chemistry

Specialty Chemistry

Date 2022 Jul 19

PMID 35852812

Authors

Carina Lemke

Adela Jilkova

Dominic Ferber

Annett Braune

Anja On

Patrick Johe

Alena Zikova

Tanja Schirmeister

Michael Mares

Martin Horn

Michael Gutschow

Affiliations

Soon will be listed here.

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 /K value of 37,000 M s . 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.

Citing Articles

Combination Therapy and Phytochemical-Loaded Nanosytems for the Treatment of Neglected Tropical Diseases.

Soto-Sanchez J, Garza-Trevino G Pharmaceutics. 2024; 16(10).

PMID: 39458571 PMC: 11510106. DOI: 10.3390/pharmaceutics16101239.

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

Lemke C, Jilkova A, Ferber D, Braune A, On A, Johe P Chemistry. 2022; 28(62):e202201636.

PMID: 35852812 PMC: 9826439. DOI: 10.1002/chem.202201636.

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