title: Photoreactive stapled peptides to identify and characterize BCL-2 family interaction sites by mass spectrometry.
abstract: Protein interactions dictate a myriad of cellular activities that maintain health or cause disease. Dissecting these binding partnerships, and especially their sites of interaction, fuels the discovery of signaling pathways, disease mechanisms, and next-generation therapeutics. We previously applied all-hydrocarbon peptide stapling to chemically restore α-helical shape to bioactive motifs that become unfolded when taken out of context from native signaling proteins. For example, we developed stabilized alpha-helices of BCL-2 domains (SAHBs) to dissect and target protein interactions of the BCL-2 family, a critical network that regulates the apoptotic pathway. SAHBs are α-helical surrogates that bind both stable and transient physiologic interactors and have effectively uncovered novel sites of BCL-2 family protein interaction. To leverage stapled peptides for proteomic discovery, we describe our conversion of SAHBs into photoreactive agents that irreversibly capture their protein targets and facilitate rapid identification of the peptide helix binding sites. We envision that the development of photoreactive stapled peptides will accelerate the discovery of novel and unanticipated protein interactions and how they impact health and disease.
authors: Department of Pediatric Oncology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA; Department of Pediatrics, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA.
authors: Susan
authors: S
authors: Lee
authors: Department of Pediatric Oncology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA; Department of Pediatrics, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA; Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Craig R
authors: CR
authors: Braun
authors: Department of Pediatric Oncology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA; Department of Pediatrics, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA.
authors: Gregory H
authors: GH
authors: Bird
authors: Department of Pediatric Oncology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA; Department of Pediatrics, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA. Electronic address: loren_walensky@dfci.harvard.edu.
authors: Loren D
authors: LD
authors: Walensky
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001665:Binding Sites; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000378:metabolism*; D017319:Photosensitizing Agents / Q000032:analysis / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D040901:Proteomics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis / Q000378:metabolism*
chemical_list: D010455:Peptides; D017319:Photosensitizing Agents; D019253:Proto-Oncogene Proteins c-bcl-2
keywords: Apoptosis; BAD; BAK; BAX; BCL-2 family; BCL-XL; BH3; BID; BIM; Binding site; Glucokinase; MCL-1; Mass spectrometry; PUMA; Photoaffinity labeling; Photoreactive; Protein interaction; Stabilized Alpha-Helix (SAH); Stabilized Alpha-Helix of BCL-2 domain (SAHB); Stapled peptide