Intracelluar delivery of A20 protein inhibits TNFα-induced NF-κB activation

Protein Expr Purif. 2018 Mar:143:14-19. doi: 10.1016/j.pep.2017.10.005. Epub 2017 Oct 4.

Abstract

A20 (also known as TNFAIP3) is a potent anti-inflammatory protein that suppresses many intracellular signaling pathways induced by inflammatory cytokines and bacterial and viral pathogens. The anti-inflammatory function of A20 depends on its modulation of or binding to polyubiquitin chains on key signaling proteins in the nuclear factor-κB (NF-κB) pathway. To test whether A20 can be used as therapeutic agent in these inflammatory diseases, we prepared a recombinant cell-penetrating form of A20 (TAT-A20) for intracellular delivery and examined its effect on tumor necrosis factor-α (TNFα)-induced NF-κB activation. We observed that TAT-A20 was effectively transduced into cells within 30 min, whereas A20 protein without TAT motive was not. TAT-A20 also inhibited NF-κB induction in fibroblasts stimulated with TNFα. These results suggest that increasing intracellular level of A20 can be an effective means to suppress NF-κB activation and treat inflammatory diseases.

Keywords: A20 (TNFAIP3); Inflammation; Intracellular delivery; NF-κB; Recombinant cell-penetrating protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Penetrating Peptides / chemistry
  • Cell-Penetrating Peptides / genetics
  • Cell-Penetrating Peptides / metabolism*
  • Escherichia coli / genetics
  • HEK293 Cells
  • Humans
  • Intracellular Space / metabolism
  • NF-kappa B / antagonists & inhibitors*
  • NF-kappa B / metabolism*
  • Protein Refolding
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Tumor Necrosis Factor alpha-Induced Protein 3 / chemistry
  • Tumor Necrosis Factor alpha-Induced Protein 3 / genetics
  • Tumor Necrosis Factor alpha-Induced Protein 3 / metabolism*
  • Tumor Necrosis Factor-alpha / metabolism*

Substances

  • Cell-Penetrating Peptides
  • NF-kappa B
  • Recombinant Fusion Proteins
  • TNF protein, human
  • Tumor Necrosis Factor-alpha
  • TNFAIP3 protein, human
  • Tumor Necrosis Factor alpha-Induced Protein 3