In vivo localisation and stability of human Mcl‐1 using green fluorescent protein (GFP) fusion proteins

FEBS Letters

Edwards, Steven W¹  Akgul, Cahit¹  Moulding, Dale A¹  White, M.R.H¹ 

[1] School of Biological Sciences, Life Sciences Building, University of Liverpool, Liverpool L69 7ZB, UK

关键词: U-937;    Apoptosis;    Bcl-2;    Fluorescence imaging;    Mitochondrion;   

DOI  :  10.1016/S0014-5793(00)01809-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Mcl-1 is an anti-apoptotic member of the Bcl-2 family of proteins. We have expressed full length and mutated GFP:Mcl-1 fusion proteins to define structural motifs that control protein localisation and stability. When expressed in U-937 cells, full length Mcl-1 locates primarily within mitochondria and its half-life was approximately 3 h, which was identical to the native, endogenously expressed protein. When the terminal 20 amino acids from the C-terminus of the protein were detected, the protein was diffused in the cytoplasm, but its stability was unaffected. This confirms that this region is responsible for efficient targeting to mitochondria. Surprisingly, deletion of 104 amino acids (residues 79–183) that contain putative PEST sequences and other stability regulating motifs, did not affect protein stability.

【 授权许可】

Unknown