Effect of ultrasound on DNA polymerase reactions: monitoring on a 27-MHz quartz crystal microbalance.
Effects of ultrasound irradiation on DNA polymerase (Klenow fragment, KF) reactions were studied on the template/primer DNA-immobilized quartz crystal microbalance (QCM). Under ultrasound irradiation, binding of KF to the DNA was suppressed due to the decrease of the binding rate constant (k(1)) and the increase of the dissociation rate constant (k(-)(1)). The catalytic elongation rate (k(cat)) was increased, but the stability of the KF/DNA/monomer ternary complex (K(m)) was decreased by the ultrasound irradiation. Ultrasound effects are discussed in correlation with the conformation changes of domain structures in KF.
Hoshino
Yu
Y
Department of Biomolecular Engineering and Frontier Collaborative Research Center, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
Kawasaki
Takayoshi
T
Okahata
Yoshio
Y
D001483:Base Sequence; D001704:Biopolymers / Q000737:chemistry; D003460:Crystallization; D004247:DNA / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000528:radiation effects; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009991:Oscillometry; D011485:Protein Binding; D011791:Quartz; D014465:Ultrasonics
D001704:Biopolymers; D046911:Macromolecular Substances; D011791:Quartz; D004247:DNA; D004259:DNA-Directed DNA Polymerase