Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis

Paul C. Trippier, Kevin Tianmeng Zhao, Susan G. Fox, Isaac T. Schiefer, Radhia Benmohamed, Jason Moran, Donald R. Kirsch, Richard I. Morimoto, Richard B. Silverman

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

(Chemical Equation Presented) Amyotrophic lateral sclerosis (ALS) is a progressive and ultimately fatal neurodegenerative disease. Pyrazolone containing small molecules have shown significant disease attenuating efficacy in cellular and murine models of ALS. Pyrazolone based affinity probes were synthesized to identify high affi nity binding partners and ascertain a potential biological mode of action. Probes were confirmed to be neuroprotective in PC12-SOD1G93A cells. PC12-SOD1G93A cell lysates were used for protein pull-down, affinity purification, and subsequent proteomic analysis using LC-MS/MS. Proteomics identified the 26S proteasome regulatory subunit 4 (PSMC1), 26S proteasome regulatory subunit 6B (PSMC4), and T-complex protein 1 (TCP-1) as putative protein targets. Coincubation with appropriate competitors confirmed the authenticity of the proteomics results. Activation of the proteasome by pyrazolones was demonstrated in the absence of exogenous proteasome inhibitor and by restoration of cellular protein degradation of a fluorogenic proteasome substrate in PC12-SOD1G93A cells. Importantly, supplementary studies indicated that these molecules do not induce a heat shock response. We propose that pyrazolones represent a rare class of molecules that enhance proteasomal activation in the absence of a heat shock response and may have therapeutic potential in ALS.

Original languageEnglish (US)
Pages (from-to)823-829
Number of pages7
JournalACS Chemical Neuroscience
Volume5
Issue number9
DOIs
StatePublished - Sep 17 2014

Fingerprint

PC12 Cells
Amyotrophic Lateral Sclerosis
Proteasome Endopeptidase Complex
Pyrazolones
Proteomics
Heat-Shock Response
Chemical activation
Molecules
Proteins
Proteasome Inhibitors
Neurodegenerative diseases
Fluorescent Dyes
Neurodegenerative Diseases
Proteolysis
Therapeutics
Restoration
Purification
Degradation
pyrazolone
Substrates

Keywords

  • Amyotrophic lateral sclerosis
  • Drug discovery
  • Neurodegeneration
  • Proteasome activator
  • Pyrazolone
  • Target identification

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Cognitive Neuroscience
  • Cell Biology

Cite this

Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis. / Trippier, Paul C.; Zhao, Kevin Tianmeng; Fox, Susan G.; Schiefer, Isaac T.; Benmohamed, Radhia; Moran, Jason; Kirsch, Donald R.; Morimoto, Richard I.; Silverman, Richard B.

In: ACS Chemical Neuroscience, Vol. 5, No. 9, 17.09.2014, p. 823-829.

Research output: Contribution to journalArticle

Trippier, PC, Zhao, KT, Fox, SG, Schiefer, IT, Benmohamed, R, Moran, J, Kirsch, DR, Morimoto, RI & Silverman, RB 2014, 'Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis', ACS Chemical Neuroscience, vol. 5, no. 9, pp. 823-829. https://doi.org/10.1021/cn500147v
Trippier, Paul C. ; Zhao, Kevin Tianmeng ; Fox, Susan G. ; Schiefer, Isaac T. ; Benmohamed, Radhia ; Moran, Jason ; Kirsch, Donald R. ; Morimoto, Richard I. ; Silverman, Richard B. / Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis. In: ACS Chemical Neuroscience. 2014 ; Vol. 5, No. 9. pp. 823-829.
@article{e1ff925b4f8f4094a22b54ea950cbb26,
title = "Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis",
abstract = "(Chemical Equation Presented) Amyotrophic lateral sclerosis (ALS) is a progressive and ultimately fatal neurodegenerative disease. Pyrazolone containing small molecules have shown significant disease attenuating efficacy in cellular and murine models of ALS. Pyrazolone based affinity probes were synthesized to identify high affi nity binding partners and ascertain a potential biological mode of action. Probes were confirmed to be neuroprotective in PC12-SOD1G93A cells. PC12-SOD1G93A cell lysates were used for protein pull-down, affinity purification, and subsequent proteomic analysis using LC-MS/MS. Proteomics identified the 26S proteasome regulatory subunit 4 (PSMC1), 26S proteasome regulatory subunit 6B (PSMC4), and T-complex protein 1 (TCP-1) as putative protein targets. Coincubation with appropriate competitors confirmed the authenticity of the proteomics results. Activation of the proteasome by pyrazolones was demonstrated in the absence of exogenous proteasome inhibitor and by restoration of cellular protein degradation of a fluorogenic proteasome substrate in PC12-SOD1G93A cells. Importantly, supplementary studies indicated that these molecules do not induce a heat shock response. We propose that pyrazolones represent a rare class of molecules that enhance proteasomal activation in the absence of a heat shock response and may have therapeutic potential in ALS.",
keywords = "Amyotrophic lateral sclerosis, Drug discovery, Neurodegeneration, Proteasome activator, Pyrazolone, Target identification",
author = "Trippier, {Paul C.} and Zhao, {Kevin Tianmeng} and Fox, {Susan G.} and Schiefer, {Isaac T.} and Radhia Benmohamed and Jason Moran and Kirsch, {Donald R.} and Morimoto, {Richard I.} and Silverman, {Richard B.}",
year = "2014",
month = "9",
day = "17",
doi = "10.1021/cn500147v",
language = "English (US)",
volume = "5",
pages = "823--829",
journal = "ACS Chemical Neuroscience",
issn = "1948-7193",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Proteasome activation is a mechanism for pyrazolone small molecules displaying therapeutic potential in amyotrophic lateral sclerosis

AU - Trippier, Paul C.

AU - Zhao, Kevin Tianmeng

AU - Fox, Susan G.

AU - Schiefer, Isaac T.

AU - Benmohamed, Radhia

AU - Moran, Jason

AU - Kirsch, Donald R.

AU - Morimoto, Richard I.

AU - Silverman, Richard B.

PY - 2014/9/17

Y1 - 2014/9/17

N2 - (Chemical Equation Presented) Amyotrophic lateral sclerosis (ALS) is a progressive and ultimately fatal neurodegenerative disease. Pyrazolone containing small molecules have shown significant disease attenuating efficacy in cellular and murine models of ALS. Pyrazolone based affinity probes were synthesized to identify high affi nity binding partners and ascertain a potential biological mode of action. Probes were confirmed to be neuroprotective in PC12-SOD1G93A cells. PC12-SOD1G93A cell lysates were used for protein pull-down, affinity purification, and subsequent proteomic analysis using LC-MS/MS. Proteomics identified the 26S proteasome regulatory subunit 4 (PSMC1), 26S proteasome regulatory subunit 6B (PSMC4), and T-complex protein 1 (TCP-1) as putative protein targets. Coincubation with appropriate competitors confirmed the authenticity of the proteomics results. Activation of the proteasome by pyrazolones was demonstrated in the absence of exogenous proteasome inhibitor and by restoration of cellular protein degradation of a fluorogenic proteasome substrate in PC12-SOD1G93A cells. Importantly, supplementary studies indicated that these molecules do not induce a heat shock response. We propose that pyrazolones represent a rare class of molecules that enhance proteasomal activation in the absence of a heat shock response and may have therapeutic potential in ALS.

AB - (Chemical Equation Presented) Amyotrophic lateral sclerosis (ALS) is a progressive and ultimately fatal neurodegenerative disease. Pyrazolone containing small molecules have shown significant disease attenuating efficacy in cellular and murine models of ALS. Pyrazolone based affinity probes were synthesized to identify high affi nity binding partners and ascertain a potential biological mode of action. Probes were confirmed to be neuroprotective in PC12-SOD1G93A cells. PC12-SOD1G93A cell lysates were used for protein pull-down, affinity purification, and subsequent proteomic analysis using LC-MS/MS. Proteomics identified the 26S proteasome regulatory subunit 4 (PSMC1), 26S proteasome regulatory subunit 6B (PSMC4), and T-complex protein 1 (TCP-1) as putative protein targets. Coincubation with appropriate competitors confirmed the authenticity of the proteomics results. Activation of the proteasome by pyrazolones was demonstrated in the absence of exogenous proteasome inhibitor and by restoration of cellular protein degradation of a fluorogenic proteasome substrate in PC12-SOD1G93A cells. Importantly, supplementary studies indicated that these molecules do not induce a heat shock response. We propose that pyrazolones represent a rare class of molecules that enhance proteasomal activation in the absence of a heat shock response and may have therapeutic potential in ALS.

KW - Amyotrophic lateral sclerosis

KW - Drug discovery

KW - Neurodegeneration

KW - Proteasome activator

KW - Pyrazolone

KW - Target identification

UR - http://www.scopus.com/inward/record.url?scp=84924980848&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84924980848&partnerID=8YFLogxK

U2 - 10.1021/cn500147v

DO - 10.1021/cn500147v

M3 - Article

C2 - 25001311

AN - SCOPUS:84924980848

VL - 5

SP - 823

EP - 829

JO - ACS Chemical Neuroscience

JF - ACS Chemical Neuroscience

SN - 1948-7193

IS - 9

ER -