Cooperative, non-specific binding of a zinc finger peptide to DNA.
The DNA binding and structural properties of Xfin-31 (Lee, M.S., Gippert, G.P., Soman, K.V., Case, D.A. and Wright, P.E., 1989, Science 245, 635-637), a twenty five amino acid zinc finger peptide, in the reduced, oxidized and zinc complex forms, as well as the fourteen residue helical segment of the zinc finger (residues 12-25) have been compared using affinity coelectrophoresis (ACE) and circular dichroism (CD) spectroscopy. The zinc complex and oxidized peptides bind cooperatively to DNA although the cooperativity factor, omega, is more than 15-fold greater for the zinc complex. The reduced peptide in the absence of zinc and the helical segment do not bind cooperatively (omega = 1). Hence, the binding constant for singly contiguous sites (K omega) ranges over 100-fold for the various peptides even though the intrinsic binding constants (K) are similar. An increase in binding order and affinity for the other forms of Xfin-31 is correlated with an increasing similarity of the CD spectrum to that of the Xfin-31 zinc complex. The surprising DNA binding activity of the oxidized peptide may result from hydrophobic interactions between the amino-terminal loop formed by the Cys3-Cys6 disulfide bond and conserved hydrophobic residues in the carboxyl-terminal segment. Xfin-31 may be a particularly useful model for studying several poorly understood aspects of cooperative, non-specific DNA binding since it is small, has a stable, well-defined structure, and structures of zinc fingers bound to DNA have been determined.
Nedved
M L
ML
Department of Chemistry and Biochemistry, University of Delaware, Newark 19716.
Moe
G R
GR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D007700:Kinetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010455:Peptides; D011089:Polydeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins; D015032:Zinc / Q000378:metabolism; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D010455:Peptides; D011089:Polydeoxyribonucleotides; D011994:Recombinant Proteins; C072294:zinc finger peptide Xfin-31; D004247:DNA; D015032:Zinc