Optimising histidine rich peptides for efficient DNA delivery in the presence of serum.
We recently showed that the antibacterial histidine rich amphipathic peptide LAH4 has significant DNA transfection capabilities in the absence of serum. To further understand the transfection process and to develop the peptides for future applications, we have combined a range of biochemical and biophysical techniques, including fluorescence assisted cell sorting and (2)H solid-state NMR, to characterise the initial binding of the peptide/DNA complexes to the cell surface and the subsequent release of the complexes from the endosome in the presence of serum. Our results show that both primary and secondary peptide structure play important roles in both of these processes. Specifically, we show that an ideal helix length and positioning of the histidine residues should be maintained to obtain optimal resistance to serum effects and release of DNA from the endosome. Inclusion of d-amino acids at the peptide termini does not reduce serum effects however further enrichment of the peptides with histidine residues can enhance transfection efficiency in the presence of serum. The detailed understanding of these two key stages in the transfection process shows that LAH4-L1 and its derivatives are likely to be highly efficient and robust vectors for a range of applications.
Mason
A James
AJ
Faculté de chimie, Université Louis Pasteur/CNRS LC3-UMR7177, Institut le Bel, 4 rue Blaise Pascal, F-67070 Strasbourg, France. jmason@chimie.u-strasbg.fr
Leborgne
Christian
C
Moulay
Gilles
G
Martinez
Amélie
A
Danos
Olivier
O
Bechinger
Burkhard
B
Kichler
Antoine
A
D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002784:Cholesterol / Q000737:chemistry; D004247:DNA; D004337:Drug Carriers; D005434:Flow Cytometry; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008156:Luciferases / Q000378:metabolism; D008956:Models, Chemical; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D044967:Serum / Q000737:chemistry*; D014162:Transfection
D004337:Drug Carriers; D008055:Lipids; D010455:Peptides; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; C050294:1-palmitoyl-2-oleoylglycero-3-phosphoserine; D006639:Histidine; D004247:DNA; D002784:Cholesterol; D008156:Luciferases; C028694:1-palmitoyl-2-oleoylphosphatidylcholine