Proteasome-associated cysteine deubiquitinases are molecular targets of environmental optical brightener compounds

J Cell Biochem. 2019 Aug;120(8):14065-14075. doi: 10.1002/jcb.28682. Epub 2019 Apr 8.

Abstract

The levels of organic pollutants, such as optical brightener (OB) compounds, in the global environment have been increasing in recent years. The toxicological effects and signal transduction systems associated with OB toxicity have not been thoroughly studied. The ubiquitin-proteasome system (UPS) plays a crucial role in regulating multiple essential cellular processes, and proteasome-associated cysteine deubiquitinases (DUBs), ubiquitin C-terminal hydrolase L5 (UCHL5) and USP14, are two major regulators for (de)ubiquitination and stability of many important target proteins. Therefore, potential inhibition of UCHL5 and USP14 activities by some environmental chemicals might cause in vivo toxicity. In the current study we hypothesize that electrophilic OB compounds, such as 4,4'-diamino-2,2'-stilbenedisulfonic acid(DAST), fluorescent brightener 28 (FB-28) and FB-71, can interact with the catalytic triads (CYS, HIS, and ASP) of UCHL5 and USP14 and inhibit their enzymatic activities, leading to cell growth suppression. This hypothesis is supported by our findings presented in this study. Results from in silico computational docking and ubiquitin vinyl sulfone assay confirmed the UCHL5/USP14-inhibitory activities of these OB compounds that have potencies in an order of: FB-71 > FB-28 > DAST. Furthermore, inhibition of these two proteasomal DUBs by OBs resulted in cell growth inhibition and apoptosis induction in two human breast cancer cell models. In addition, we found that OB-mediated DUB inhibition triggers a feedback reaction in which expression of UCHL5 and USP14 proteins is increased to compromise the suppressed activities. Our study suggests that these commonly used OB compounds may target and inhibit proteasomal cysteine DUBs, which should contribute to their toxicological effects in vivo.

Keywords: degradation; environmental pollutant; fluorescent brightening agent; molecular target; posttranslational modification.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Catalytic Domain
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cysteine / metabolism*
  • Deubiquitinating Enzymes / metabolism*
  • Environmental Pollutants / chemistry
  • Environmental Pollutants / toxicity*
  • Humans
  • Ligands
  • Molecular Docking Simulation
  • Poly(ADP-ribose) Polymerases / metabolism
  • Proteasome Endopeptidase Complex / metabolism*
  • Ubiquitin Thiolesterase / antagonists & inhibitors
  • Ubiquitin Thiolesterase / metabolism

Substances

  • Environmental Pollutants
  • Ligands
  • USP14 protein, human
  • Poly(ADP-ribose) Polymerases
  • Deubiquitinating Enzymes
  • UCHL5 protein, human
  • Ubiquitin Thiolesterase
  • Proteasome Endopeptidase Complex
  • Cysteine