Motif-programmed artificial protein induces apoptosis in several cancer cells by disrupting mitochondria.
By combinatorially assembling two natural motifs, respectively, associated with protein transduction (PTD) and induction of apoptosis (BH3), we previously synthesized an artificial protein (#284) that is taken up into cells, where it induces apoptosis. Here we used cluster analysis of GI(50) (average concentration required for 50% growth inhibition), as well as immunohistochemical and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling analyses to further characterize the capacity of #284 to induce apoptosis in a panel of 39 cancer cell lines. Our results showed that #284 preferentially inhibited the growth of several cancer cells with a GI(50) of approximately 5 microM, which is in the range of conventional anticancer drugs such as cisplatin and etoposide. In breast cancer HBC-4 cells, #284 caused mitochondrial aggregation and induced apoptosis in a BH3 motif-dependent manner. Moreover, transfection of the artificial gene that encodes #284 led to effective expression of the artificial protein within cells, which in turn caused apoptosis at a level similar to that seen in naturally occurring apoptosis inducers, Noxa/Bax transfectants. These findings suggest that synthetic proteins created by reprogramming peptide motifs have the potential to serve as novel agents useful in the treatment of cancer.
Saito
Hirohide
H
Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Koto-ku, Tokyo, Japan.
Minamisawa
Tamiko
T
Yamori
Takao
T
Shiba
Kiyotaka
K
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000633:toxicity; D017209:Apoptosis; D045744:Cell Line, Tumor; D006801:Humans; D020287:In Situ Nick-End Labeling; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015202:Protein Engineering; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000633:toxicity; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000633:toxicity
D000970:Antineoplastic Agents; D011506:Proteins; D011993:Recombinant Fusion Proteins; C528189:artificial protein 284