N-terminal acylation of the SV40 nuclear localization signal peptide enhances its oligonucleotide binding and membrane translocation efficiencies.
Octanoyl and palmitoyl groups were coupled to the N-terminus of an analog of the SV40 nuclear localization signal peptide, SV126-133(Ser128), to study the effect of the fatty acid chain length on the complex formation with a single-stranded antisense oligodeoxynucleotide (ODN) and on the cellular uptake of the complex. The strongest binding affinity was observed for the palmitoylated peptide, indicating the better accessibility of the positively charged lysyl and arginyl side-chains to the phosphate groups due to the turn structures stabilized by the palmitoyl group. On increase of the peptide to ODN molar ratio (rM), gradual unstacking of the bases was observed, the maximal rate being reached at rM=10. At rM>10 restacking of the nucleotide bases was detected and the ODN was completely encapsulated in a liposome-like structure made up of palmitoylated peptides. Cell translocation experiments revealed a highly efficient cell transport of the ODN by palmitoylated SV40 peptide at rM>10.
Laczkó
Ilona
I
Institute of Biophysics, Biological Research Center, Szeged, Hungary. laczko@nucleus.szbk.u-szeged.hu
Váró
György
G
Bottka
Sándor
S
Bálint
Zoltán
Z
Illyés
Eszter
E
Vass
Elemér
E
Bertrand
Jean-Remy
JR
Malvy
Claude
C
Hollósi
Miklós
M
D000215:Acylation; D000818:Animals; D001692:Biological Transport; D002210:Caprylates / Q000737:chemistry*; D002463:Cell Membrane Permeability; D002942:Circular Dichroism; D004337:Drug Carriers; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D018625:Microscopy, Atomic Force; D008968:Molecular Conformation; D041681:NIH 3T3 Cells; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000378:metabolism* / Q000494:pharmacology; D009842:Oligopeptides / Q000737:chemistry*; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D010169:Palmitic Acids / Q000737:chemistry*; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics; D034802:RNA-Binding Protein EWS; D017550:Spectroscopy, Fourier Transform Infrared
D002210:Caprylates; D004337:Drug Carriers; C106031:EWS-FLI fusion protein; D020319:Oligodeoxyribonucleotides, Antisense; D009842:Oligopeptides; D015514:Oncogene Proteins, Fusion; D010169:Palmitic Acids; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; C402304:SV40 nuclear localization peptide; D049452:Green Fluorescent Proteins