2. Academic Validation
  3. Antagonists of anaphase-promoting complex (APC)-2-cell cycle and apoptosis regulatory protein (CARP)-1 interaction are novel regulators of cell growth and apoptosis

Antagonists of anaphase-promoting complex (APC)-2-cell cycle and apoptosis regulatory protein (CARP)-1 interaction are novel regulators of cell growth and apoptosis

  • J Biol Chem. 2011 Nov 4;286(44):38000-38017. doi: 10.1074/jbc.M111.222398.
Vineshkumar Thidil Puliyappadamba 1,  Wenjuan Wu 1,  Debra Bevis 2,  Liyue Zhang 3,  Lisa Polin 4,  Robert Kilkuskie 2,  Russell L Finley Jr 5,  Scott D Larsen 6,  Edi Levi 7,  Fred R Miller 8,  Anil Wali 9,  Arun K Rishi 10
Affiliations

Affiliations

  • 1 Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 2 Michigan High-throughput Screening Center, Kalamazoo Valley Community College, Kalamazoo, Michigan 49003.
  • 3 Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan 48201; John D. Dingell Veterans Affairs Medical Center, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 4 Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan 48201; Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 5 Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 6 College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109.
  • 7 John D. Dingell Veterans Affairs Medical Center, Wayne State University School of Medicine, Detroit, Michigan 48201; Department of Pathology, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 8 Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan 48201; Department of Pathology, Wayne State University School of Medicine, Detroit, Michigan 48201; Breast Cancer Program, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 9 John D. Dingell Veterans Affairs Medical Center, Wayne State University School of Medicine, Detroit, Michigan 48201; Department of Surgery, Wayne State University School of Medicine, Detroit, Michigan 48201.
  • 10 Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan 48201; John D. Dingell Veterans Affairs Medical Center, Wayne State University School of Medicine, Detroit, Michigan 48201; Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan 48201; Breast Cancer Program, Wayne State University School of Medicine, Detroit, Michigan 48201. Electronic address: Rishia@Karmanos.org.
PMID: 21903591 DOI: 10.1074/jbc.M111.222398
Abstract

CARP-1/CCAR1, a perinuclear phosphoprotein, is a regulator of cell growth and Apoptosis signaling. Although CARP-1 is a regulator of chemotherapy-dependent Apoptosis, it is also a part of the NF-κB proteome and a co-activator of steroid/thyroid nuclear receptors as well as β-catenin signaling. Our yeast two-hybrid screen revealed CARP-1 binding with the anaphase-promoting complex/cyclosome E3 ubiquitin Ligase component APC-2 protein. CARP-1 also binds with anaphase-promoting complex/cyclosome co-activators Cdc20 and Cdh1. Following mapping of the minimal epitopes involved in CARP-1 binding with APC-2, a fluorescence polarization assay was established that indicated a dissociation constant (K(d)) of 480 nm for CARP-1/APC-2 binding. Fluorescence polarization assay-based high throughput screening of a chemical library yielded several small molecule antagonists of CARP-1/APC-2 binding, termed CARP-1 functional mimetics. CFM-4 (1(2-chlorobenzyl)-5'-phenyl-3'H-spiro[indoline-3,2'-[1,3,4]thiadiazol]-2-one), a lead compound, binds with and stimulates CARP-1 expression. CFM-4 prevents CARP-1 binding with APC-2, causes G(2)M cell cycle arrest, and induces Apoptosis with an IC(50) range of 10-15 μm. Apoptosis signaling by CFM-4 involves activation of Caspase-8 and -9 and caspase-mediated ubiquitin-proteasome pathway-independent loss of cyclin B1 and Cdc20 proteins. Depletion of CARP-1, however, interferes with CFM-4-dependent cell growth inhibition, activation of caspases, and Apoptosis. Because CFM-4 also suppresses growth of drug-resistant human breast Cancer cells without affecting the growth of human breast epithelial MCF-10A cells, elevating CARP-1 by CFM-4 and consequent Apoptosis could in principle be exploited to further elucidate, and perhaps effectively target, often deregulated cell cycle pathways in pathological conditions, including Cancer.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-103255
    98.85%, CARP-1/APC-2 Binding Antagonist
  • HY-116473
    CARP-1/APC-2 Binding Antagonist