FRET-Based Cathepsin Probes for Simultaneous Detection of Cathepsin B and D Activities

Zhengxiao Zhang; Eiji Nakata; Yuya Shibano; Takashi Morii

doi:10.1002/cbic.202200319

FRET-Based Cathepsin Probes for Simultaneous Detection of Cathepsin B and D Activities

Chembiochem. 2022 Oct 6;23(19):e202200319. doi: 10.1002/cbic.202200319. Epub 2022 Aug 31.

Authors

Zhengxiao Zhang¹, Eiji Nakata¹, Yuya Shibano¹, Takashi Morii¹

Affiliation

¹ Institute of Advanced Energy, Kyoto University, Uji, Kyoto, 611-0011, Japan.

PMID: 35929606
DOI: 10.1002/cbic.202200319

Abstract

Fluorescent cathepsin probes were prepared by modification of peptidic substrates for cathepsin B (CTSB) and cathepsin D (CTSD) with FRET pairs. Fluorophores with distinguishable emission characteristics were applied to CTSB and CTSD probes with their appropriate quenchers to simultaneously monitor the activity of CTSB and/or CTSD. Conjugation of both the CTSB and CTSD probes with short single-stranded DNA drastically increased their reactivity to cathepsins over the parent probes possibly by improving their solubility. The activity of CTSB and CTSD were simultaneously detected by using these orthogonal FRET-based cathepsin probes.

Keywords: DNA-peptide conjugation; cathepsin; fluorescent probe; multi detection system.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cathepsin B* / genetics
Cathepsin B* / metabolism
Cathepsin D* / genetics
Cathepsin D* / metabolism
DNA, Single-Stranded
Fluorescence Resonance Energy Transfer

Substances

DNA, Single-Stranded
Cathepsin B
Cathepsin D