TY - JOUR

T1 - Complexes of DNA with cationic peptides

T2 - Conditions of formation and factors effecting internalization by mammalian cells

AU - Dizhe, E. B.

AU - Ignatovich, I. A.

AU - Burov, S. V.

AU - Pohvoscheva, A. V.

AU - Akifiev, B. N.

AU - Efremov, A. M.

AU - Perevozchikov, A. P.

AU - Orlov, S. V.

N1 - Funding Information: This work was supported by the Russian Foundation for Basic Research (grants 04 04 48683 and 06 04 49784).

PY - 2006/12

Y1 - 2006/12

N2 - 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.

AB - 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.

KW - Cationic peptides

KW - Endocytosis

KW - K8

KW - Non-viral gene transfer approaches

KW - Tat peptide

UR - http://www.scopus.com/inward/record.url?scp=33845629764&partnerID=8YFLogxK

U2 - 10.1134/S0006297906120108

DO - 10.1134/S0006297906120108

M3 - Article

C2 - 17223788

AN - SCOPUS:33845629764

VL - 71

SP - 1350

EP - 1356

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

SN - 0006-2979

IS - 12

ER -