Public reporting burden for this collection of information is estimated to average 1 hour per response, induding the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Washington Headquarters Services, Directorate for Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302, and The Bcl-2 family proteins are key regulators of programmed cell death, in health and major human diseases, including cancer. Their pro-or anti-apoptotic functions are regulated by subcellular location, as the proteins cycle between soluble and membrane-bound forms; by dimerization with other Bcl-2 family members; by binding to other non-homologous proteins; and by formation of membrane pores that are believed to regulate apoptosis by perturbing mitochondrial physiology. The solution structures of several Bcl-2 family proteins are very similar despite their antagonistic activities, however, the structures of the membrane-associated proteins are not known and may be key to their opposing functions. The goals of this project are: (1) to determine the structures of the meribrane-asaociated Bd-2 proteins; and (2) to determine their mechanism of apoptosis regulation. The research stategy combines NMR structure determination in lipid environments with biological assays carried out in parallel with structure determination.
INTRODUCTIONThe goal of this project was to determine the structures of the Bcl-2 family proteins in their membraneassociated forms. The Bcl-2 family proteins play major regulatory roles in programmed cell death, or apoptosis, and exert their activities through dimerization with other Bcl-2 family members, binding to nonhomologous proteins, and the formation of membrane pores, believed to regulate apoptosis by perturbing mitochondrial physiology. Their functions are also regulated by subcellular location as the proteins cycle between soluble and membrane-bound forms. The soluble structures of several pro-and anti-apoptotic Bcl-2 family proteins are very similar, despite their antagonistic activities, however, the structures of their membrane-associated forms have not been previously examined, and may be important for understanding their opposing functions. Moreover, most of the previous structural and functional studies have focused on soluble truncated proteins, lacking the C-terminal 20-residue hydrophobic domain, which is present in many of the family members and is important for membrane targeting.The support of this grant has enabled us to initiate a major research effort to determine the structures of membrane-bound and full-length Bcl-2 proteins. The results from these studies serve as a platform for additional structural and biological experiments aimed at examining the structural consequences of modificat...