Target protein interactions of indole-3-carbinol and the highly potent derivative 1-benzyl-I3C with the C-terminal domain of human elastase uncouples cell cycle arrest from apoptotic signaling.

Abstract

Elastase is the only currently identified target protein for indole-3-carbinol (I3C), a naturally occurring hydrolysis product of glucobrassicin in cruciferous vegetables such as broccoli, cabbage, and Brussels sprouts that induces a cell cycle arrest and apoptosis of human breast cancer cells. In vitro elastase enzymatic assays demonstrated that I3C and at lower concentrations its more potent derivative 1-benzyl-indole-3-carbinol (1-benzyl-I3C) act as non-competitive allosteric inhibitors of elastase activity. Consistent with these results, in silico computational simulations have revealed the first predicted interactions of I3C and 1-benzyl-I3C with the crystal structure of human neutrophil elastase, and identified a potential binding cluster on an external surface of the protease outside of the catalytic site that implicates elastase as a target protein for both indolecarbinol compounds. The Δ205 carboxyterminal truncation of elastase, which disrupts the predicted indolecarbinol binding site, is enzymatically active and generates a novel I3C resistant enzyme. Expression of the wild type and Δ205 elastase in MDA-MB-231 human breast cancer cells demonstrated that the carboxyterminal domain of elastase is required for the I3C and 1-benzyl-I3C inhibition of enzymatic activity, accumulation of the unprocessed form of the CD40 elastase substrate (a tumor necrosis factor receptor family member), disruption of NFκB nuclear localization and transcriptional activity, and induction of a G1 cell cycle arrest. Surprisingly, expression of the Δ205 elastase molecule failed to reverse indolecarbinol stimulated apoptosis, establishing an elastase-dependent bifurcation point in anti-proliferative signaling that uncouples the cell cycle and apoptotic responses in human breast cancer cells.

• Authors: Ida Aronchik, Tony Chen, Kathleen A Durkin, Marshall S Horwitz, Maria N Preobrazhenskaya, Leonard F Bjeldanes, Gary L Firestone
• Journal: Molecular carcinogenesis (Mol Carcinog) Vol. 51 Issue 11 Pg. 881-94 (Nov 2012) ISSN: 1098-2744 [Electronic] United States
• PMID: 22012859 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2011 Wiley Periodicals, Inc.

Chemical References

• Enzyme Inhibitors
• Indoles
• NF-kappa B
• indole-3-carbinol
• Leukocyte Elastase

Topics

• Apoptosis (drug effects)
• Breast Neoplasms (drug therapy, enzymology, genetics, metabolism)
• Cell Cycle (drug effects)
• Cell Line, Tumor
• Enzyme Inhibitors (chemistry, pharmacology)
• Female
• Humans
• Indoles (chemistry, pharmacology)
• Leukocyte Elastase (antagonists & inhibitors, chemistry, genetics, metabolism)
• Models, Molecular
• Mutation
• NF-kappa B (analysis, metabolism)
• Protein Structure, Tertiary
• Signal Transduction (drug effects)
• Vegetables (chemistry)