A Fluorescence-Polarization-Based Lipopolysaccharide-Caspase-4 Interaction Assay for the Development of Inhibitors.
Jinsu AnSo Yeon KimEun Gyeong YangHak Suk ChungPublished in: Molecules (Basel, Switzerland) (2022)
Recognition of intracellular lipopolysaccharide (LPS) by Caspase-4 (Casp-4) is critical for host defense against Gram-negative pathogens. LPS binds to the N-terminal caspase activation and recruitment domain (CARD) of procaspase-4, leading to auto-proteolytic activation followed by pro-inflammatory cytokine release and pyroptotic cell death. Aberrant hyper-activation of Casp-4 leads to amplification of the inflammatory response linked to sepsis. While the active site of a caspase has been targeted with peptide inhibitors, inhibition of LPS-Casp-4 interaction is an emerging strategy for the development of selective inhibitors with a new mode of action for treating infectious diseases and sepsis induced by LPS. In this study, a high-throughput screening (HTS) system based on fluorescence polarization (FP) was devised to identify inhibitors of the LPS and Casp-4 interaction. Using HTS and IC 50 determination and subsequently showing inhibited Casp-4 activity, we demonstrated that the LPS-Casp-4 interaction is a druggable target for Casp-4 inhibition and possibly a non-canonical inflammatory pathway.
Keyphrases
- inflammatory response
- cell death
- gram negative
- lps induced
- lipopolysaccharide induced
- toll like receptor
- anti inflammatory
- multidrug resistant
- infectious diseases
- intensive care unit
- acute kidney injury
- cell cycle arrest
- oxidative stress
- single molecule
- immune response
- septic shock
- reactive oxygen species
- cancer therapy
- signaling pathway
- drug delivery
- mass spectrometry
- nucleic acid