Biophysical characterization of the DNA binding and condensing properties of adenoviral core peptide μ (mu)

被引:51
作者
Keller, M [1 ]
Tagawa, T [1 ]
Preuss, M [1 ]
Miller, AD [1 ]
机构
[1] Univ London Imperial Coll Sci Technol & Med, Dept Chem, Genet Therapeut Ctr, London SW7 2AZ, England
关键词
D O I
10.1021/bi0156299
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cationic peptides containing Lys and Arg residues interact with DNA via charge-charge interactions and are known to play an important role in DNA charge neutralization and condensation processes. In this paper, we describe investigations of the interaction of the cationic adenovirus core complex peptide p (mu) with a dodecameric ODN (12 bp) and pDNA (7528 bp) using a combination of fluorescence spectroscopy, circular dichroism spectroscopy, isothermal titration calorimetry, and photon correlation spectroscopy. Comparisons are made with protamine. a cationic peptide well-known for DNA charge neutralization and condensation. Equilibrium dissociation constants are derived independently by both CD and ITC methods for the interaction between protamine or mu with pDNA (K-d = 0.6 - 1 muM). Thermodynamic data are also obtained by ITC, indicating strong charge-charge interactions. The interaction of protamine with pDNA takes place with decreasing entropy (-28.7 cal mol(-1) K-1), Unusually, the interaction of mu with pDNA takes place with increasing entropy (DeltaSdegrees(bind) = 11.3 cal mol(-1) K-1). Although protamine and mu appear to destabilize pDNA double helix character to similar extents, according. to CD thermal titration analyses, PCS studies show that interactions between MU and pDNA result in the formation of significantly more size-stable condensed particles than protamine. The enhanced flexibility and size stability of mu-DNA (MD) particles (80-110 nm) compared to protamine counterparts Suggest that MD particles are ideal for use as a part of new nonviral gene delivery vectors.
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页码:652 / 659
页数:8
相关论文
共 51 条
[1]   CHARACTERIZATION OF THE ADENOVIRUS-2 VIRION PROTEIN, MU [J].
ANDERSON, CW ;
YOUNG, ME ;
FLINT, SJ .
VIROLOGY, 1989, 172 (02) :506-512
[2]   CONDENSATION OF DNA BY TRIVALENT CATIONS .1. EFFECTS OF DNA LENGTH AND TOPOLOGY ON THE SIZE AND SHAPE OF CONDENSED PARTICLES [J].
ARSCOTT, PG ;
LI, AZ ;
BLOOMFIELD, VA .
BIOPOLYMERS, 1990, 30 (5-6) :619-630
[3]   SPECIFIC RECOGNITION OF APURINIC SITES IN DNA BY A TRYPTOPHAN-CONTAINING PEPTIDE [J].
BEHMOARAS, T ;
TOULME, JJ ;
HELENE, C .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1981, 78 (02) :926-930
[4]   A TRYPTOPHAN-CONTAINING PEPTIDE RECOGNIZES AND CLEAVES DNA AT APURINIC SITES [J].
BEHMOARAS, T ;
TOULME, JJ ;
HELENE, C .
NATURE, 1981, 292 (5826) :858-859
[5]   Physical stability and in-vitro gene expression efficiency of nebulised lipid-peptide-DNA complexes [J].
Birchall, JC ;
Kellaway, IW ;
Gumbleton, M .
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 2000, 197 (1-2) :221-231
[6]   CONDENSATION OF DNA BY MULTIVALENT CATIONS - CONSIDERATIONS ON MECHANISM [J].
BLOOMFIELD, VA .
BIOPOLYMERS, 1991, 31 (13) :1471-1481
[7]   DNA condensation [J].
Bloomfield, VA .
CURRENT OPINION IN STRUCTURAL BIOLOGY, 1996, 6 (03) :334-341
[8]  
Boulikas T, 1997, INT J ONCOL, V10, P317
[9]   H-1-NMR STUDY OF THE BASE-PAIRING REACTIONS OF D(GGAATTCC) - SALT EFFECTS ON THE EQUILIBRIA AND KINETICS OF STRAND ASSOCIATION [J].
BRAUNLIN, WH ;
BLOOMFIELD, VA .
BIOCHEMISTRY, 1991, 30 (03) :754-758
[10]   INTERACTIONS OF AROMATIC RESIDUES OF PROTEINS WITH NUCLEIC-ACIDS - FLUORESCENCE STUDIES OF BINDING OF OLIGOPEPTIDES CONTAINING TRYPTOPHAN AND TYROSINE RESIDUES TO POLYNUCLEOTIDES [J].
BRUN, F ;
TOULME, JJ ;
HELENE, C .
BIOCHEMISTRY, 1975, 14 (03) :558-563