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Literature summary for 2.7.7.7 extracted from

  • Yasukawa, K.; Konishi, A.; Shinomura, M.; Nagaoka, E.; Fujiwara, S.
    Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases (2012), Biochem. Biophys. Res. Commun., 427, 654-658.
    View publication on PubMed
Search for more literature for 2.7.7.7

Cloned(Commentary)

Cloned (Comment) Organism
expression of soluble mutant L329A in Escherichia coli strain BL21(DE3) codon plus Thermotoga petrophila
N-terminally His6-tagged M1 DNA polymerase in Eschrichia coli strain BL21(DE3) codon plus Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
L329A site-directed mutagenesis Thermotoga petrophila

Inhibitors

Inhibitors Comment Organism Structure
additional information high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases Thermotoga petrophila
additional information high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases Thermus thermophilus
poly(rA)-p(dT)45 inhibitory effect towards the reverse transcriptase activity of K4polL329A Thermotoga petrophila
poly(rA)-p(dT)45 inhibitory effect towards the reverse transcriptase activity of M1pol Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information the reaction rate of K4polL329A exhibits a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)45 concentrations Thermotoga petrophila
additional information
-
 additional information the reaction rate of M1pol exhibits a saturated profile of the Michaelis-Menten kinetics for dTTP concentrations but a substrate inhibition profile for poly(rA)-p(dT)45 concentrations Thermus thermophilus
0.23
-
 dTTP pH 8.2, 50°C, recombinant M1pol Thermus thermophilus
0.28
-
 dTTP pH 8.2, 50°C, recombinant enzyme mutant K4polL329A Thermotoga petrophila

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
deoxynucleoside triphosphate + DNAn Thermotoga petrophila
-
 diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn Thermus thermophilus
-
 diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn Thermus thermophilus M1
-
 diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn Thermotoga petrophila K4
-
 diphosphate + DNAn+1
-
 ?
additional information Thermotoga petrophila only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 ?
-
 ?
additional information Thermotoga petrophila K4 only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Thermotoga petrophila
-
-
-
Thermotoga petrophila K4
-
-
-
Thermus thermophilus K4Q1U9 isolated from a hot spring at Kagoshima, Japan
-
Thermus thermophilus M1 K4Q1U9 isolated from a hot spring at Kagoshima, Japan
-

Purification (Commentary)

Purification (Comment) Organism
N-terminally His6-tagged M1 DNA polymerase from Eschrichia coli strain BL21(DE3) codon plus by nickel affinity chromatography and dialysis Thermus thermophilus
recombinant soluble mutant L329A from Escherichia coli strain BL21(DE3) codon plus partially by heat treatment at 85°C for 30 min Thermotoga petrophila

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxynucleoside triphosphate + DNAn
-
 Thermotoga petrophila diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn
-
 Thermus thermophilus diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn DNA-dependent DNA polymerase activity to incorporate dNTP into gapped M13mp2 DNA Thermus thermophilus diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn
-
 Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn DNA-dependent DNA polymerase activity to incorporate dNTP into gapped M13mp2 DNA Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
deoxynucleoside triphosphate + DNAn
-
 Thermotoga petrophila K4 diphosphate + DNAn+1
-
 ?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermotoga petrophila diphosphate + DNAn+1
-
 ?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermus thermophilus diphosphate + DNAn+1
-
 ?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermus thermophilus M1 diphosphate + DNAn+1
-
 ?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)45 Thermotoga petrophila K4 diphosphate + DNAn+1
-
 ?
additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 Thermotoga petrophila ?
-
 ?
additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, no activity with the wild-type enzyme Thermotoga petrophila ?
-
 ?
additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, EC 2.7..7.49 Thermotoga petrophila K4 ?
-
 ?
additional information only the enzyme mutant T326A/L324A/Q384A/F388A/m4008A/Y438A shows RNA-dependent DNA polymerase activity, no activity with the wild-type enzyme Thermotoga petrophila K4 ?
-
 ?

Synonyms

Synonyms Comment Organism
K4pol
-
 Thermotoga petrophila
M1 DNA polymerase
-
 Thermus thermophilus
M1pol
-
 Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
 assay at Thermotoga petrophila
50
-
 assay at Thermus thermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
37 65 optimum above Thermotoga petrophila

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1
-
 dTTP pH 8.2, 50°C, recombinant M1pol Thermus thermophilus
1.7
-
 dTTP pH 8.2, 50°C, recombinant enzyme mutant K4polL329A Thermotoga petrophila

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
 assay at Thermotoga petrophila
8.2
-
 assay at Thermus thermophilus

General Information

General Information Comment Organism
evolution the enzyme belongs to the DNA polymerase family A Thermotoga petrophila
evolution the enzyme belongs to the DNA polymerase family A Thermus thermophilus
additional information nine residues, Tyr326, Leu329, Gln384, Lys387, Phe388, Met408, Asn422, Tyr438, and Phe451, are predicted to be involved in DNA/RNA distinction Thermotoga petrophila