title: Mutagenesis of the positively charged conserved residues in the 5' exonuclease domain of Taq DNA polymerase.
abstract: Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction method. Taq DNA polymerase has a domain at the amino terminus (residue 1 to 290) that has a 5' exonuclease activity and a domain at the C-terminus that catalyzes polymerase reaction. Taq DNA polymerase is classified into the pol I family which is represented by E. coli DNA polymerase I. The alignment of amino acid sequences for the 5' exonuclease domains of the pol I family DNA polymerases shows six highly conserved sequences called motifs A to F. Motif C contains three positively charged residues such as 74Arg, 82Lys and 85Arg which might be involved in catalysis. In order to understand the function of those residues, they are mutagenized to alanine. The 5' exonucleolytic activities of those mutated 5' exonucleases decreased by 80 to 90%, thereby implying that three positively charged residues play certain roles in the 5' exonuclease catalysis.
authors.affiliation: Korea Research Institute of Bioscience and Biotechnology, Taejon.
authors.forename: Y
authors.initials: Y
authors.lastname: Kim
authors.forename: J S
authors.initials: JS
authors.lastname: Kim
authors.forename: Y
authors.initials: Y
authors.lastname: Park
authors.forename: C S
authors.initials: CS
authors.lastname: Chang
authors.forename: S W
authors.initials: SW
authors.lastname: Suh
authors.forename: D S
authors.initials: DS
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000502:physiology; D001665:Binding Sites / Q000235:genetics; D017124:Conserved Sequence; D005090:Exodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D019914:Taq Polymerase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D000596:Amino Acids; D019914:Taq Polymerase; D005090:Exodeoxyribonucleases