Identification of small molecule inhibitors of the functional in vitro reconstituted apoptosome

摘要

Apoptosis(programmed cell death)is an important proeess in a wide variety of biological systems,including normal cell turnover,the immune system and embryonic development.Inappropriate apoptosis is involved in many human pathologies,including neurodegenerative diseases such as Alzheimer's and Huntington's,ischaemia.autoimmune disorders and several forms of cancer.Apoptotic stimuli induces signaling cascades that converge into a common pathway,the activation of the caspases,a family of cysteine aspartyl proteases.Because of the fatal consequences of apopotosis malfunctioning,the activation of caspases is scrupulously controlled.Such level of control is ensured in certain apotototic pathways by the participation of cell-organells as the mitochondria.The mitochondria-mediated or intrinsic pathway of proteolytic maturation of dormant caspases,responds to most apoptotic signals and is triggered by the liberation to the cytosol of proapoptotic proteins from the mitochondrial inter-membrane space,in particular cytochrome c and Smac/Diablo.The formation of the maeromoleeular complex named apoptosome is one of the most relevant events of this pathway.The apoptosome is a 700 kDa holoenzyme multiprotein complex formed by cytochrome c-activaled Apaf-1(apoptotic protease-activating factor),ATP and various copies of procaspase-9.In this maeromoleeular complex apoptosome-associated caspase-9 is activated and can then activate effector caspases.Thus,to contribute to the identification of molecules that could aminoriate dissease-associaled apoptosis we wanted to develop inhibitors of the activity of the apoptosome.