title: [Design and synthesis of peptides capable of specific binding to DNA].
abstract: In the present communication, design, synthesis and DNA binding activities of the following two peptides are reported: Dns-Gly-Ala-Gln-Lys-Leu-Ala-Cly-Lys-Val-Gly-Thr-Lys-Val-Lys-Val-Gl y-Thr-Lys-Thr - Val-OH (I) and [(H-Ala-Lys-Leu-Ala-Thr-Lys-Ala-Gly-Val-Lys-Gln-Gln-Ser-Ile-Gln-Leu-Ile- Thr- Ala-Aca-Lys-Aca)2Lys-Aca]2Lys-Val-OH (II), where Aca = NH(CH2)5CO--; Dns is a residue of 5-dimethylaminonaphtalene-1-sulfonic acid. Peptide I contains a large fraction (ca.30%) of valyl and threonyl residues, which possess a high potential for beta structure formation. Peptide II contains four repeats of the amino acid sequence present in the presumed DNA binding helix-turn-helix unit of 434 Cro repressor. These four domains are linked in such a way that two domains can interact with two halves a 14 base pair long operator site on DNA. From CD studies we have found that peptide I is in a random coil conformation in the aqueous solution in the presence of 20% trifluoroethanol. By contrast, amino acid residues of peptide II assume alpha helical, beta and random coiled conformations under the same conditions. A change in the secondary structure of the two peptides upon binding to DNA is observed. The difference CD spectra obtained by subtracting the spectra of free DNA from the spectra of peptide I--DNA complexes gives rise to a beta-like pattern. The difference CD spectra obtained for complexes of peptide II with various natural and synthetic DNAs suggest that alpha-beta-transition takes place in the presumed helix-turn-helix repeat units of peptide II upon binding to DNA. Peptide I binds more strongly to poly(dG).poly(dC) than to poly(dA).poly(dT) and poly[d(GC)].poly[d(GC)]. The binding takes place in the minor DNA groove because minor groove binding antibiotic sibiromycin can displace peptide I from a complex with poly(dG).poly(dC). Analysis of footprinting diagramms shows that peptide I specifically protects phosphodiester bonds within operator sites OR1 and OR2 of phage lambda from nuclease cleavage. By contrast, peptide II does not react specifically with operators OR1, OR2 and OR3 of phage 434 although it forms very tight complexes with DNA which are stable in the presence of 1M NH4F.
authors.forename: S L
authors.initials: SL
authors.lastname: Grokhovskiĭ
authors.forename: A N
authors.initials: AN
authors.lastname: Surovaia
authors.forename: N Iu
authors.initials: NIu
authors.lastname: Sidorova
authors.forename: H
authors.initials: H
authors.lastname: Votavova
authors.forename: J
authors.initials: J
authors.lastname: Sponar
mesh_terms: D000818:Animals; D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D004268:DNA-Binding Proteins / Q000138:chemical synthesis* / Q000378:metabolism; D003850:Deoxyribonuclease I; D009690:Nucleic Acid Conformation; D010455:Peptides / Q000138:chemical synthesis* / Q000378:metabolism; D011089:Polydeoxyribonucleotides / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D010455:Peptides; D011089:Polydeoxyribonucleotides; D004247:DNA; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I
vernacular_title: Konstruirovanie i sintez peptidov, sposobnykh spetsificheski sviazyvat'sia s DNK.