Inactivation of DNA polymerase by adenosine 2',3'-riboepoxide 5'-triphosphate allows estimation of the primers affinity.
Template-primer dependent inactivation of human DNA polymerase alpha and Klenow fragment of E. coli DNA polymerase I by adenosine 2',3'-riboepoxide 5'-triphosphate was used for quantitative analysis of the Kd values for oligonucleotide primers of different length. The Kd values are smaller by a factor of 2.5 than the Km values for the same primers determined in the reaction of DNA polymerization in the case of DNA polymerase alpha. The Kd and Km values are nearly the same for Klenow fragment. Such approach to the determination of Km/Kd ratio can likely be used for detailed quantitative analysis of DNA polymerases.
Podust
V N
VN
Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, Novosibirsk.
Korobeinicheva
T O
TO
Nevinsky
G A
GA
Levina
A S
AS
Lavrik
O I
OI
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D004256:DNA Polymerase I / Q000032:analysis / Q000037:antagonists & inhibitors*; D004258:DNA Polymerase III / Q000032:analysis / Q000037:antagonists & inhibitors*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology; D007700:Kinetics; D009841:Oligonucleotides / Q000032:analysis*; D011061:Poly A; D011071:Poly T; D011072:Poly U; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D018894:Reverse Transcriptase Inhibitors; D013223:Statistics as Topic
D009841:Oligonucleotides; D018894:Reverse Transcriptase Inhibitors; D011061:Poly A; D011071:Poly T; D011072:Poly U; C016205:adenosine 2',3'-riboepoxide 5'-triphosphate; D000255:Adenosine Triphosphate; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; D004258:DNA Polymerase III