Proton equilibria in the minor groove of DNA.
Poisson-Boltzmann calculations by Pack and co-workers suggest the presence of regions of increased hydrogen ion density in the grooves of DNA. As an experimental test of this prediction, we have attached proton-sensitive probes, with variable linker lengths, to random-sequence DNA at G sites in the minor groove. The amino groups of beta-alanine, gamma-aminobutyric acid (GABA), and epsilon-aminocaproic acid have been coupled at pH 5, via a formaldehyde link, to the exocyclic amino group of guanine, utilizing a reaction that has been extensively investigated by Hanlon and co-workers. The resulting adducts at pH 5 retained duplex B form but exhibited typical circular dichroism (CD) changes previously shown to be correlated with the presence of a net positive charge in the minor groove. Increases in the solvent pH reversed the CD spectral changes in a manner suggesting deprotonation of the carboxylic acid group of the adduct. These data were used to calculate an apparent pK(a) for the COOH. The pK(a) was increased by 2.4 units for beta-alanine, by 1.7 units for GABA, and by 1.5 units for epsilon-amino caproic acid, relative to their values in the free amino acid. This agrees well with Poisson-Boltzmann calculations and the energy minimization of the structures of the adducts that place the carboxyl groups in acidic domains whose hydrogen ion density is approximately 2 orders of magnitude greater than that of bulk solvent.
Hanlon
S
S
Department of Biochemistry, University of Illinois College of Medicine, Chicago 60612, USA.
Wong
L
L
Pack
G R
GR
D000409:Alanine; D000596:Amino Acids; D002208:Caproates; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000138:chemical synthesis / Q000737:chemistry*; D005998:Glycine; D007700:Kinetics; D008958:Models, Molecular; D008962:Models, Theoretical; D009690:Nucleic Acid Conformation; D013816:Thermodynamics; D005680:gamma-Aminobutyric Acid
D000596:Amino Acids; D002208:Caproates; D018736:DNA Adducts; D005680:gamma-Aminobutyric Acid; D004247:DNA; D000409:Alanine; D005998:Glycine