Synthesis and biophysical characterization of stabilized alpha-helices of BCL-2 domains.
Rational design of compounds to mimic the functional domains of BCL-2 family proteins requires chemical reproduction of the biologic complexity afforded by the relatively large and folded surfaces of BCL-2 homology (BH) domain peptide alpha-helices. Because the intermolecular handshakes of BCL-2 proteins are so critical to controlling cellular fate, we undertook the development of a toolbox of peptidic ligands that harness the natural potency and specificity of BH alpha-helices to interrogate and potentially medicate the deregulated apoptotic pathways of human disease. To overcome the classic deficiencies of peptide reagents, including loss of bioactive structure in solution, rapid proteolytic degradation in vivo, and cellular impermeability, we developed a new class of compounds based on hydrocarbon stapling of BH3 death domain peptides. Here we describe the chemical synthesis of Stabilized Alpha-Helices of BCL-2 domains or SAHBs, and the analytical methods used to characterize their secondary structure, proteolytic stability, and cellular penetrance.
Bird
Gregory H
GH
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.
Bernal
Federico
F
Pitter
Kenneth
K
Walensky
Loren D
LD
D051023:BH3 Interacting Domain Death Agonist Protein / Q000138:chemical synthesis; D002463:Cell Membrane Permeability; D002918:Chymotrypsin / Q000378:metabolism; D002942:Circular Dichroism; D015195:Drug Design; D005434:Flow Cytometry; D018613:Microscopy, Confocal; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000138:chemical synthesis*; D014357:Trypsin / Q000378:metabolism
D051023:BH3 Interacting Domain Death Agonist Protein; D019253:Proto-Oncogene Proteins c-bcl-2; D002918:Chymotrypsin; D014357:Trypsin