RRC ID 39301
Author Niidome T, Takaji K, Urakawa M, Ohmori N, Wada A, Hirayama T, Aoyagi H.
Title Chain length of cationic alpha-helical peptide sufficient for gene delivery into cells.
Journal Bioconjug. Chem.
Abstract To define the minimal peptide length needed for gene delivery into mammalian cells, we synthesized several peptides with shortened chain lengths from the amino-termini of the original amphiphilic peptides (4(6), Ac-LARL-LARL-LARL-LRAL-LRAL-LRAL-NH( 2,) and Hel 11-7, KLLK-LLLK-LWKK-LLKL-LK), which have been known to have gene transfer abilities into cells. Each synthetic peptide was studied for its ability to bind and aggregate with plasmid DNA and the structural change of the peptide caused by binding with the DNA to establish a relative in vitro gene transfection efficiency in COS-7 cells. As a result, the deletion of eight amino acid residues of 4(6) had little influence on their ability, whereas that of 12 amino acid residues remarkably reduced the abilities to make aggregates and transfer the DNA into the cell. In the case of the Hel 11-7 series peptides, deletion of amino acid residues caused a considerable reduction in abilities to bind and form aggregates with DNA and to transfer the DNA into cell in due order. In summary, 16 and 17 amino acid residues were sufficient to form aggregates with the DNA and transfer the DNA into the cells in the deletion series of 4(6) and Hel 11-7, respectively. Furthermore, it was indicated that reduction of membrane perturbation activity of the peptide-DNA complex due to deletion of the peptide chain length caused suppression of the transfection efficiency even if the complex was incorporated into the cells. Transfer of the complex to cytosol mediated by membrane perturbation activity of the peptide is an important step for efficient protein expression from its cDNA. The results of this study will make it easy to design and synthesize a functional gene carrier molecule such as a carbohydrate-modified peptide used in targeted gene delivery.
Volume 10(5)
Pages 773-80
Published 1999-9
DOI 10.1021/bc990012d
PII bc990012d
PMID 10502342
MeSH Amino Acid Sequence Animals CHO Cells COS Cells Cations Cell Membrane / metabolism Circular Dichroism Cricetinae DNA / administration & dosage* DNA / chemistry DNA / metabolism Gene Transfer Techniques* HeLa Cells Humans Molecular Sequence Data Particle Size Peptides / administration & dosage* Peptides / chemical synthesis Peptides / chemistry* Peptides / metabolism Plasmids / genetics Protein Structure, Secondary Structure-Activity Relationship Tumor Cells, Cultured
IF 4.485
Times Cited 38
WOS Category CHEMISTRY, ORGANIC CHEMISTRY, MULTIDISCIPLINARY BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells