Complementary protocols to evaluate inhibitors against the DnaK chaperone network

Aweon Richards, Gideon K. Yawson, Brock Nelson, Tania J. Lupoli

Research output: Contribution to journalArticlepeer-review

Abstract

Bacterial DnaK belongs to the Hsp70 chaperone family, which plays a critical role in maintaining proteostasis by catalyzing protein folding, and is a proposed antibacterial target in the pathogen Mycobacterium tuberculosis. Here, we describe an experimental toolbox for evaluating inhibitors against the mycobacterial DnaK chaperone network: a coupled-enzymatic assay to monitor ATPase activity, a proteolytic cleavage assay to study DnaK conformational changes upon ligand addition, as well as a protein renaturation assay to assess chaperone function. For complete details on the use and execution of this protocol, please refer to Hosfelt et al. (2021).

Original languageEnglish (US)
Article number101381
JournalSTAR Protocols
Volume3
Issue number2
DOIs
StatePublished - Jun 17 2022

Keywords

  • Molecular/Chemical Probes
  • Protein Biochemistry

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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