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Application of immobilized ATP to the study of NLRP inflammasomes.

Publication ,  Journal Article
Liao, K-C; Sandall, CF; Carlson, DA; Ulke-Lemée, A; Platnich, JM; Hughes, PF; Muruve, DA; Haystead, TAJ; MacDonald, JA
Published in: Arch Biochem Biophys
July 30, 2019

The NLRP proteins are a subfamily of the NOD-like receptor (NLR) innate immune sensors that possess an ATP-binding NACHT domain. As the most well studied member, NLRP3 can initiate the assembly process of a multiprotein complex, termed the inflammasome, upon detection of a wide range of microbial products and endogenous danger signals and results in the activation of pro-caspase-1, a cysteine protease that regulates multiple host defense pathways including cytokine maturation. Dysregulated NLRP3 activation contributes to inflammation and the pathogenesis of several chronic diseases, and the ATP-binding properties of NLRPs are thought to be critical for inflammasome activation. In light of this, we examined the utility of immobilized ATP matrices in the study of NLRP inflammasomes. Using NLRP3 as the prototypical member of the family, P-linked ATP Sepharose was determined to be a highly-effective capture agent. In subsequent examinations, P-linked ATP Sepharose was used as an enrichment tool to enable the effective profiling of NLRP3-biomarker signatures with selected reaction monitoring-mass spectrometry (SRM-MS). Finally, ATP Sepharose was used in combination with a fluorescence-linked enzyme chemoproteomic strategy (FLECS) screen to identify potential competitive inhibitors of NLRP3. The identification of a novel benzo[d]imidazol-2-one inhibitor that specifically targets the ATP-binding and hydrolysis properties of the NLRP3 protein implies that ATP Sepharose and FLECS could be applied other NLRPs as well.

Duke Scholars

Published In

Arch Biochem Biophys

DOI

EISSN

1096-0384

Publication Date

July 30, 2019

Volume

670

Start / End Page

104 / 115

Location

United States

Related Subject Headings

  • Ubiquitination
  • Protein Processing, Post-Translational
  • Phosphorylation
  • NLR Proteins
  • Inflammasomes
  • Humans
  • HEK293 Cells
  • Biochemistry & Molecular Biology
  • Adenosine Triphosphate
  • 3101 Biochemistry and cell biology
 

Citation

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ICMJE
MLA
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Liao, K.-C., Sandall, C. F., Carlson, D. A., Ulke-Lemée, A., Platnich, J. M., Hughes, P. F., … MacDonald, J. A. (2019). Application of immobilized ATP to the study of NLRP inflammasomes. Arch Biochem Biophys, 670, 104–115. https://doi.org/10.1016/j.abb.2018.12.031
Liao, Kuo-Chieh, Christina F. Sandall, David A. Carlson, Annegret Ulke-Lemée, Jaye M. Platnich, Philip F. Hughes, Daniel A. Muruve, Timothy A. J. Haystead, and Justin A. MacDonald. “Application of immobilized ATP to the study of NLRP inflammasomes.Arch Biochem Biophys 670 (July 30, 2019): 104–15. https://doi.org/10.1016/j.abb.2018.12.031.
Liao K-C, Sandall CF, Carlson DA, Ulke-Lemée A, Platnich JM, Hughes PF, et al. Application of immobilized ATP to the study of NLRP inflammasomes. Arch Biochem Biophys. 2019 Jul 30;670:104–15.
Liao, Kuo-Chieh, et al. “Application of immobilized ATP to the study of NLRP inflammasomes.Arch Biochem Biophys, vol. 670, July 2019, pp. 104–15. Pubmed, doi:10.1016/j.abb.2018.12.031.
Liao K-C, Sandall CF, Carlson DA, Ulke-Lemée A, Platnich JM, Hughes PF, Muruve DA, Haystead TAJ, MacDonald JA. Application of immobilized ATP to the study of NLRP inflammasomes. Arch Biochem Biophys. 2019 Jul 30;670:104–115.
Journal cover image

Published In

Arch Biochem Biophys

DOI

EISSN

1096-0384

Publication Date

July 30, 2019

Volume

670

Start / End Page

104 / 115

Location

United States

Related Subject Headings

  • Ubiquitination
  • Protein Processing, Post-Translational
  • Phosphorylation
  • NLR Proteins
  • Inflammasomes
  • Humans
  • HEK293 Cells
  • Biochemistry & Molecular Biology
  • Adenosine Triphosphate
  • 3101 Biochemistry and cell biology