Abstract | BACKGROUND: METHODOLOGY AND PRINCIPAL FINDINGS: Here we developed in vitro binding and cell-based functional assays to demonstrate that a nitrofuran-containing (NFC) group in EerI is the functional domain responsible for the cytotoxicity. Using both SPR and pull down assays, we show that EerI directly binds the p97 ATPase, an essential component of the ERAD machinery, via the NFC domain. An aromatic domain in EerI, although not required for p97 interaction, can localize EerI to the ER membrane, which improves its target specificity. Substitution of the aromatic module with another benzene-containing domain that maintains membrane localization generates a structurally distinct compound that nonetheless has similar biologic activities as EerI. CONCLUSIONS AND SIGNIFICANCE: Our findings reveal a class of bifunctional chemical agents that can preferentially inhibit membrane-bound p97 to disrupt ER homeostasis and to induce tumor cell death. These results also suggest that the AAA ATPase p97 may be a potential drug target for cancer therapeutics.
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Authors | Qiuyan Wang, Bidhan A Shinkre, Jin-gu Lee, Marc A Weniger, Yanfen Liu, Weiping Chen, Adrian Wiestner, William C Trenkle, Yihong Ye |
Journal | PloS one
(PLoS One)
Vol. 5
Issue 11
Pg. e15479
(Nov 12 2010)
ISSN: 1932-6203 [Electronic] United States |
PMID | 21124757
(Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)
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Chemical References |
- 1-(4-chlorophenyl)-3-(3-(4-chlorophenyl)-5,5-dimethyl-1-(3-(5-nitrofuran-2-yl)allyldienehydrazinocarbonylmethyl)-2-oxoimidazolidin-4-yl)-1-hydroxyurea
- Hydrazones
- Nuclear Proteins
- Adenosine Triphosphatases
- p97 ATPase
- Hydroxyurea
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Topics |
- Adenosine Triphosphatases
(antagonists & inhibitors, metabolism)
- Cluster Analysis
- Endoplasmic Reticulum
(metabolism)
- Gene Expression Profiling
- HEK293 Cells
- HeLa Cells
- Humans
- Hydrazones
(chemistry, metabolism, pharmacology)
- Hydroxyurea
(analogs & derivatives, chemistry, metabolism, pharmacology)
- Intracellular Membranes
(metabolism)
- Molecular Structure
- Nuclear Proteins
(antagonists & inhibitors, metabolism)
- Oligonucleotide Array Sequence Analysis
- Protein Binding
- Protein Transport
(drug effects)
- Reverse Transcriptase Polymerase Chain Reaction
- Surface Plasmon Resonance
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