Full-length Fas-associated Death Domain Protein Interacts with Short Form of Cellular FLICE Inhibitory Protein

Bulletin of the Korean chemical society

Mi Suk Jeong¹  Se Bok Jang¹ 

关键词: Overexpression;    Purification;    Full-length FADD;    c-FLIPS (short form);    Interaction;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

Fas-associated death domain protein (FADD) recruits and activates procaspase-8 through interactions between the death effector domains of these two proteins. Cellular FLICE-inhibitory protein (c-FLIP) was identified as a molecule with sequence homology to caspase-8. It has been postulated that c-FLIP prevents formation of the competent death-inducing signaling complex in a ligand-dependent manner, through its interaction with FADD and/or caspase-8. However, the interaction of FADD and c-FLIP_S (short form) in apoptosis signaling has been controversially discussed. We show the purification and the characterization of human full-length FADD and c-FLIPS expressed in Escherichia coli. The purified FADD and c-FLIP_S are shown as homogeneity, respectively, in SDS-PAGE analysis and light-scattering measurements. The folding properties of the �?-helical structure of FADD and the super-secondary structure of c-FLIP_S proteins were characterized by circular dichroism spectroscopy. Furthermore, we report here a series of biochemical and biophysical data for FADDc- FLIP_S binding in vitro. The binding of both FADD and c-FLIP_S proteins was detected by BIAcore biosensor, fluorescence measurement, and size-exclusion column (SEC).

【 授权许可】

Unknown