2Institute of Macromolecular Compounds, Russian Academy of Sciences, Bolshoi pr. V. O. 31, 199004 St. Petersburg, Russia; fax: (812) 328-6869; E-mail: email@example.com
3St. Petersburg State University, Universitetskaya Nab. 7/9, 199034 St. Petersburg, Russia; fax: (812) 234-0310; E-mail: firstname.lastname@example.org
* To whom correspondence should be addressed.
Received March 3, 2006; Revision received September 14, 2006
This work was devoted to the study of conditions of the formation of DNA/K8 complex and analysis of factors effecting the entry of DNA/K8 complex into mammalian cells in comparison with DNA complexes with arginine-rich fragment (47-57) of human immunodeficiency virus (type 1) transcription factor Tat (Tat peptide). The stoichiometry of positively charged DNA/K8 complexes has been studied for the first time. Non-cooperative character of DNA-K8 interaction was revealed. It has been shown that along with the positive charge of such complexes, the presence of an excess of free K8 peptide in the culture medium is a necessary condition for maximal efficiency of cell transfection with DNA/K8 complexes. A stimulatory effect of free K8 peptide on the efficiency of mammalian cell transfection by DNA/K8 complexes is likely to be mediated by the interactions of cationic peptide K8 with negatively charged proteoglycans on the cell surface, which leads to protection of DNA/K8 complexes from disruption by cellular heparan sulfates. However, the protective role of free cationic peptides depends not only on their positive charge, but also on the primary structure of the peptide. In contrast with the results obtained for DNA complexes with molecular conjugates based on poly-L-lysine, the aggregation of DNA/K8 complexes leads to a significant increase in the expression of transferred gene.
KEY WORDS: non-viral gene transfer approaches, cationic peptides, endocytosis, K8, Tat peptide