Characterization of a novel cytotoxic cell-penetrating peptide derived from p14ARF protein.
The tumor suppressor p14ARF is widely deregulated in many types of cancers and is believed to function as a failsafe mechanism, inhibiting proliferation and inducing apoptosis as cellular response to a high oncogene load. We have found that a 22-amino-acid-long peptide derived from the N-terminal part of p14ARF, denoted ARF(1-22), which has previously been shown to mimic the function of p14ARF, has cell-penetrating properties. This peptide is internalized to the same extent as the cell-penetrating peptide (CPP) TP10 and dose-dependently decreases proliferation in MCF-7 and MDA MB 231 cells. Uptake of the ARF(1-22) peptide is associated with low membrane disturbance, measured by deoxyglucose and lactate dehydrogenase (LDH) leakage, as compared to its scrambled peptide. Also, flow cytometric analysis of annexin V/propidium iodide (PI) binding and Hoechst staining of nuclei suggest that ARF(1-22) induces apoptosis, whereas scrambled or inverted peptide sequences have no effect. The ARF(1-22) peptide mainly translocates cells through endocytosis, and is found intact inside cells for at least 3 hours. To our knowledge, this is the first time a CPP having pro-apoptopic activity has been designed from a protein.
Johansson
Henrik J
HJ
Department of Neurochemistry, Stockholm University, Stockholm, Sweden. henrik@neurochem.su.se
El-Andaloussi
Samir
S
Holm
Tina
T
Mäe
Maarja
M
Jänes
Jaak
J
Maimets
Toivo
T
Langel
Ulo
U
D000595:Amino Acid Sequence; D017209:Apoptosis / Q000235:genetics; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000235:genetics; D049109:Cell Proliferation; D003603:Cytotoxins / Q000138:chemical synthesis* / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D018716:Molecular Mimicry / Q000235:genetics; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000235:genetics / Q000502:physiology; D010455:Peptides / Q000138:chemical synthesis* / Q000235:genetics / Q000502:physiology*; D025803:Tumor Suppressor Protein p14ARF / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D003603:Cytotoxins; D020135:Peptide Nucleic Acids; D010455:Peptides; D025803:Tumor Suppressor Protein p14ARF