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In vivo localisation and stability of human Mcl‐1 using green fluorescent protein (GFP) fusion proteins
Author(s) -
Akgul Cahit,
Moulding Dale A,
White M.R.H,
Edwards Steven W
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01809-3
Subject(s) - green fluorescent protein , fusion protein , cytoplasm , amino acid , biology , mitochondrion , microbiology and biotechnology , biochemistry , chemistry , gene , recombinant dna
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.