A novel branched TAT(47-57) peptide for selective Ni(2+) introduction into the human fibrosarcoma cell nucleus.
A TAT47-57 peptide was modified on the N-terminus by elongation with a 2,3-diaminopropionic acid residue and then by coupling of two histidine residues on its N-atoms. This branched peptide could bind to Ni under physiological conditions as a 1 : 1 complex. We demonstrated that the complex was quantitatively taken up by human fibrosarcoma cells, in contrast to Ni(2+) ions. Ni localization (especially at the nuclei) was confirmed by imaging using both scanning X-ray fluorescence microscopy and Newport Green fluorescence. A competitive assay with Newport Green showed that the latter displaced the peptide ligand from the Ni-complex. Ni(2+) delivered as a complex with the designed peptide induced substantially more DNA damage than when introduced as a free ion. The availability of such a construct opens up the way to investigate the importance of the nucleus as a target for the cytotoxicity, genotoxicity or carcinogenicity of Ni(2+).
CNRS/UPPA, LCABIE, UMR5254, Hélioparc, 2, av. Pr. Angot, F-64053 Pau, France. lukasz.szyrwiel@univ-pau.fr.
Łukasz
Ł
Szyrwiel
Mari
M
Shimura
Junko
J
Shirataki
Satoshi
S
Matsuyama
Akihiro
A
Matsunaga
Bartosz
B
Setner
Łukasz
Ł
Szczukowski
Zbigniew
Z
Szewczuk
Kazuto
K
Yamauchi
Wiesław
W
Malinka
Laurent
L
Chavatte
Ryszard
R
Łobinski
D000595:Amino Acid Sequence; D002273:Carcinogens / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004249:DNA Damage / Q000187:drug effects; D005354:Fibrosarcoma / Q000139:chemically induced* / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D009532:Nickel / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity*; D061848:Optical Imaging; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000378:metabolism*
D002273:Carcinogens; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; C497137:tat peptide (47-57), Human immunodeficiency virus 1; D009532:Nickel