Assays to measure small molecule Hsp70 agonist activity in vitro and in vivo

Anal Biochem. 2025 Feb:697:115712. doi: 10.1016/j.ab.2024.115712. Epub 2024 Nov 9.

Abstract

Hsp70 prevents protein aggregation and is cytoprotective, but sustained Hsp70 overexpression is problematic. Therefore, we characterized small molecule agonists that augment Hsp70 activity. Because cumbersome assays were required to assay agonists, we developed cell-based and in vivo assays in which disease-associated consequences of Hsp70 activation can be quantified. One assay uses an optogenetic system in which the formation of TDP-43 inclusions can be controlled, and the second assay employs a zebrafish model for acute kidney injury (AKI). These complementary assays will facilitate future work to identify new Hsp70 agonists as well as optimized agonist derivatives.

Keywords: Acute kidney injury (AKI); Amyotrophic lateral sclerosis (ALS); Hsp70; Molecular chaperone; Proteostasis; TDP-43.

MeSH terms

  • Animals
  • DNA-Binding Proteins / agonists
  • DNA-Binding Proteins / metabolism
  • HSP70 Heat-Shock Proteins* / agonists
  • HSP70 Heat-Shock Proteins* / metabolism
  • Humans
  • Optogenetics / methods
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology
  • Zebrafish*

Substances

  • HSP70 Heat-Shock Proteins
  • Small Molecule Libraries
  • DNA-Binding Proteins