title: Dissection of the BCL-2 family signaling network with stabilized alpha-helices of BCL-2 domains.
abstract: The BCL-2 family of apoptotic proteins regulates the critical balance between cellular life and death and, thus, has become the focus of intensive basic science inquiry and a fundamental target for therapeutic development in oncology and other diseases. Classified based on the presence of conserved alpha-helical motifs and pro- and anti-apoptotic functionalities, BCL-2 proteins participate in a complex interaction network that determines cellular fate. The identification of BCL-2 homology domain 3 (BH3) as a critical death helix that engages and regulates BCL-2 family proteins has inspired the development of molecular tools to decode and drug the interaction network. Stabilized Alpha-Helices of BCL-2 domains (SAHBs) are structurally reinforced, protease-resistant, and cell-permeable compounds that retain the specificity of native BH3 death ligands and, therefore, serve as ideal reagents to dissect BCL-2 family interactions in vitro and in vivo. Here, we describe the in vitro and cell-based methods that exploit SAHB compounds to determine the functional consequences of BH3 interactions in regulating apoptosis.
authors.affiliation: Department of Pediatric Oncology and the Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Pitter
authors.forename: Federico
authors.initials: F
authors.lastname: Bernal
authors.forename: James
authors.initials: J
authors.lastname: Labelle
authors.forename: Loren D
authors.initials: LD
authors.lastname: Walensky
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism*; D045304:Cytochromes c / Q000378:metabolism; D003911:Dextrans / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D005454:Fluorescence Polarization; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D017434:Protein Structure, Tertiary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000502:physiology; D015398:Signal Transduction; D051028:bcl-2-Associated X Protein / Q000502:physiology
chemical_list: D017304:Annexin A5; D051023:BH3 Interacting Domain Death Agonist Protein; D003911:Dextrans; D008081:Liposomes; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C015219:fluorescein isothiocyanate dextran; D045304:Cytochromes c; D016650:Fluorescein-5-isothiocyanate