A high throughput substrate binding assay reveals hexachlorophene as an inhibitor of the ER-resident HSP70 chaperone GRP78

Andrew J. Ambrose, Christopher J. Zerio, Jared Sivinski, Cody J. Schmidlin, Taoda Shi, Alison B. Ross, Kimberly J. Widrick, Steven Johnson, Donna D. Zhang, Eli Chapman

Research output: Contribution to journalArticle

Abstract

Glucose-regulated protein 78 (GRP78) is the ER resident 70 kDa heat shock protein 70 (HSP70) and has been hypothesized to be a therapeutic target for various forms of cancer due to its role in mitigating proteotoxic stress in the ER, its elevated expression in some cancers, and the correlation between high levels for GRP78 and a poor prognosis. Herein we report the development and use of a high throughput fluorescence polarization-based peptide binding assay as an initial step toward the discovery and development of GRP78 inhibitors. This assay was used in a pilot screen to discover the anti-infective agent, hexachlorophene, as an inhibitor of GRP78. Through biochemical characterization we show that hexachlorophene is a competitive inhibitor of the GRP78-peptide interaction. Biological investigations showed that this molecule induces the unfolded protein response, induces autophagy, and leads to apoptosis in a colon carcinoma cell model, which is known to be sensitive to GRP78 inhibition.

Original languageEnglish (US)
Pages (from-to)1689-1693
Number of pages5
JournalBioorganic and Medicinal Chemistry Letters
Volume29
Issue number14
DOIs
StatePublished - Jul 15 2019
Externally publishedYes

Fingerprint

Hexachlorophene
HSP70 Heat-Shock Proteins
Assays
Throughput
Substrates
Unfolded Protein Response
Fluorescence Polarization
Autophagy
Anti-Infective Agents
glucose-regulated proteins
Neoplasms
Colon
Fluorescence
Cells
Polarization
Apoptosis
Carcinoma
Peptides
Molecules

Keywords

  • Autophagy
  • Cancer
  • Chaperone
  • ER stress
  • ERAD
  • GRP78/BiP
  • HSP70
  • UPR

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

A high throughput substrate binding assay reveals hexachlorophene as an inhibitor of the ER-resident HSP70 chaperone GRP78. / Ambrose, Andrew J.; Zerio, Christopher J.; Sivinski, Jared; Schmidlin, Cody J.; Shi, Taoda; Ross, Alison B.; Widrick, Kimberly J.; Johnson, Steven; Zhang, Donna D.; Chapman, Eli.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 29, No. 14, 15.07.2019, p. 1689-1693.

Research output: Contribution to journalArticle

Ambrose, AJ, Zerio, CJ, Sivinski, J, Schmidlin, CJ, Shi, T, Ross, AB, Widrick, KJ, Johnson, S, Zhang, DD & Chapman, E 2019, 'A high throughput substrate binding assay reveals hexachlorophene as an inhibitor of the ER-resident HSP70 chaperone GRP78', Bioorganic and Medicinal Chemistry Letters, vol. 29, no. 14, pp. 1689-1693. https://doi.org/10.1016/j.bmcl.2019.05.041
Ambrose, Andrew J. ; Zerio, Christopher J. ; Sivinski, Jared ; Schmidlin, Cody J. ; Shi, Taoda ; Ross, Alison B. ; Widrick, Kimberly J. ; Johnson, Steven ; Zhang, Donna D. ; Chapman, Eli. / A high throughput substrate binding assay reveals hexachlorophene as an inhibitor of the ER-resident HSP70 chaperone GRP78. In: Bioorganic and Medicinal Chemistry Letters. 2019 ; Vol. 29, No. 14. pp. 1689-1693.
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