In situ monitoring of a trace intermediate during DNA phosphorylation by T4 polynucleotide kinase for transient kinetic studies.
Formation and decomposition of the enzyme-substrate (ES) complex during phosphorylation by T4 polynucleotide kinase (T4 PNK) of dsDNAs were monitored using a highly sensitive quartz crystal microbalance (QCM) to determine kinetic parameters, which were characterised in comparison with those of other enzymes such as DNA polymerase and exo- and endo-nucleases.
Department of Biomolecular Engineering, Tokyo Institute of Technology, B-53 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Hiroyuki
H
Furusawa
Kensuke
K
Uemura
Hiroshi
H
Yoshimine
Yoshio
Y
Okahata
D017122:Bacteriophage T4 / Q000201:enzymology*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism*; D058525:Quartz Crystal Microbalance Techniques / Q000295:instrumentation / Q000379:methods*
D004247:DNA; D011116:Polynucleotide 5'-Hydroxyl-Kinase