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Literature summary for 2.7.7.49 extracted from

  • Konishi, A.; Shinomura, M.; Yasukawa, K.
    Enzymatic characterization of human immunodeficiency virus type 1 reverse transcriptase for use in cDNA synthesis (2013), Appl. Biochem. Biotechnol., 169, 77-87.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
DMSO activation at 5-15% v/v Moloney murine leukemia virus

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged MMLV RT in Escherichia coli strain BL21(DE3) Moloney murine leukemia virus
expression of N-terminally His6-tagged HIV-1 group M and O RT subunits p51 and p66 in Escherichia coli strain BL21(DE3) Human immunodeficiency virus 1

Protein Variants

Protein Variants Comment Organism
K65R a HIV-1 O RT variant that shows increased fidelity and stability compared to the wild-type enzyme Human immunodeficiency virus 1
K65R/V75I a HIV-1 O RT variant that shows increased fidelity and stability compared to the wild-type enzyme Human immunodeficiency virus 1
R78A a HIV-1 O RT variant that shows increased fidelity and stability compared to the wild-type enzyme Human immunodeficiency virus 1

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics, overview Moloney murine leukemia virus
additional information
-
additional information Michaelis-Menten kinetics, overview Human immunodeficiency virus 1
0.0013
-
DNAn pH 8.3, 25°C, recombinant enzyme, HIV-1 O RT Human immunodeficiency virus 1
0.0014
-
DNAn pH 8.3, 25°C, recombinant enzyme, HIV-1 M RT Human immunodeficiency virus 1
0.0056
-
DNAn pH 8.3, 25°C, recombinant enzyme Moloney murine leukemia virus
0.02
-
dTTP pH 8.3, 25°C, recombinant enzyme, HIV-1 M RT Human immunodeficiency virus 1
0.03
-
dTTP pH 8.3, 25°C, recombinant enzyme, HIV-1 O RT Human immunodeficiency virus 1
0.25
-
dTTP pH 8.3, 25°C, recombinant enzyme Moloney murine leukemia virus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Moloney murine leukemia virus
Mg2+ required Human immunodeficiency virus 1

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
51000
-
1 * 66000 + 1 * 51000 or 2 * 66000 or 2 * 51000 Human immunodeficiency virus 1
66000
-
1 * 66000 + 1 * 51000 or 2 * 66000 or 2 * 51000 Human immunodeficiency virus 1

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
deoxynucleoside triphosphate + DNAn Moloney murine leukemia virus
-
diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn Human immunodeficiency virus 1
-
diphosphate + DNAn+1
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
DMSO 50% loss of activity at 15% v/v, inactivation at 25% v/v Human immunodeficiency virus 1
DMSO stable up to 15% v/v, 50% activation at 5-15% v/v, inactivation at 25% v/v Moloney murine leukemia virus
formamide 50% loss of activity at 17.5% v/v, inactivation at 15% v/v Moloney murine leukemia virus
formamide HIV-1 RT has low susceptibility to formamide, stable up to 15% v/v, inactivation at 25% v/v Human immunodeficiency virus 1

Organism

Organism UniProt Comment Textmining
Human immunodeficiency virus 1
-
group O and M virus, Group O infection is restricted to Cameroon and neighboring countries in West Central Africa, while Group M infection is spread all over the world
-
Moloney murine leukemia virus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged MMLV RT from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, dialysis, anion exchange and nickel affinity chromatography Moloney murine leukemia virus
recombinant N-terminally His6-tagged HIV-1 group M and O RT subunits p51 and p66 from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, dialysis, anion exchange and nickel affinity chromatography Human immunodeficiency virus 1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxynucleoside triphosphate + DNAn
-
Moloney murine leukemia virus diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn
-
Human immunodeficiency virus 1 diphosphate + DNAn+1
-
?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)15 Moloney murine leukemia virus diphosphate + DNAn+1
-
?
dTTP + DNAn incorporation of dTTP into poly(rA)-p(dT)15 Human immunodeficiency virus 1 diphosphate + DNAn+1
-
?
additional information HIV-1 RT has high substrate affinity and low susceptibility to formamide Moloney murine leukemia virus ?
-
?
additional information HIV-1 RT has high substrate affinity and low susceptibility to formamide, and low fidelity. Extremely high fidelity is not required for detection of a target RNA in RT-polymerase chain reaction and RNA-specific amplification, and an isothermal reaction that is widely used in clinical diagnosis, in which RT synthesizes promoter-bearing double-stranded DNA with the help of its RNase H activity Human immunodeficiency virus 1 ?
-
?

Subunits

Subunits Comment Organism
dimer
-
Moloney murine leukemia virus
dimer 1 * 66000 + 1 * 51000 or 2 * 66000 or 2 * 51000 Human immunodeficiency virus 1
More HIV-1 RT exists as a heterodimer as well as a homodimer Human immunodeficiency virus 1

Synonyms

Synonyms Comment Organism
HIV-1 M RT
-
Human immunodeficiency virus 1
HIV-1 O RT
-
Human immunodeficiency virus 1
human immunodeficiency virus type 1 reverse transcriptase
-
Human immunodeficiency virus 1
MMLV RT
-
Moloney murine leukemia virus
Moloney murine leukemia virus RT
-
Moloney murine leukemia virus
reverse transcriptase
-
Moloney murine leukemia virus
reverse transcriptase
-
Human immunodeficiency virus 1
RT
-
Moloney murine leukemia virus
RT
-
Human immunodeficiency virus 1

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25 37 assay at Moloney murine leukemia virus
25 37 assay at Human immunodeficiency virus 1

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.35
-
dTTP pH 8.3, 25°C, recombinant enzyme, HIV-1 M RT Human immunodeficiency virus 1
0.35
-
DNAn pH 8.3, 25°C, recombinant enzyme, HIV-1 M RT Human immunodeficiency virus 1
1.1
-
dTTP pH 8.3, 25°C, recombinant enzyme, HIV-1 O RT Human immunodeficiency virus 1
1.1
-
DNAn pH 8.3, 25°C, recombinant enzyme, HIV-1 O RT Human immunodeficiency virus 1
29
-
dTTP pH 8.3, 25°C, recombinant enzyme Moloney murine leukemia virus
29
-
DNAn pH 8.3, 25°C, recombinant enzyme Moloney murine leukemia virus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.3
-
assay at Moloney murine leukemia virus
8.3
-
assay at Human immunodeficiency virus 1