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Literature summary for 3.1.13.2 extracted from

  • Yasukawa, K.; Mizuno, M.; Inouye, K.
    Characterization of Moloney murine leukaemia virus/avian myeloblastosis virus chimeric reverse transcriptases (2009), J. Biochem., 145, 315-324.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Avian myeloblastosis virus
expressed in Escherichia coli Moloney murine leukemia virus

Protein Variants

Protein Variants Comment Organism
D224A two Moloney murine leukemia virus reverse transcriptase variants (named MRT-D224A and MRT-D524A) as a negative control, in which the catalytically important residue for the reverse transcription activity, Asp224 and that for the RNase H activity, Asp524, are substituted with Ala, respectively Moloney murine leukemia virus
D524A two Moloney murine leukemia virus reverse transcriptase variants (named MRT-D224A and MRT-D524A) as a negative control, in which the catalytically important residue for the reverse transcription activity, Asp224 and that for the RNase H activity, Asp524, are substituted with Ala, respectively Moloney murine leukemia virus
additional information design of four Moloney murine leukaemia virus/Avian myeloblastosis virus chimeric reverse transcriptases (named MRT-AF, MRT-AP, MRT-AT and MRT-AR) Avian myeloblastosis virus
additional information design of four Moloney murine leukemia virus/avian myeloblastosis virus chimeric reverse transcriptases (named MRT-AF, MRT-AP, MRT-AT and MRT-AR) Moloney murine leukemia virus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
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additional information Km value of Avian myeloblastosis virus reverse transcriptase (RNase H reaction) above 25 microM. KM-values for different Moloney murine leukaemia virus/Avian myeloblastosis virus chimeric reverse transcriptases with poly(rA)*p(dT)45 as a substrate: 0.013 mM for MRT-AF, 0.0070 mM for MRT-AP, 0.0089 mM for MRT-AT, and 0.0043 mM for MRT-AR Avian myeloblastosis virus
additional information
-
additional information Km value of Moloney murine leukemia virus reverse transcriptase (RNase H reaction) above 25 microM. Km values for different Moloney murine leukemia virus/avian myeloblastosis virus chimeric reverse transcriptases with poly(rA)*p(dT)45 as a substrate: 0.013 mM for MRT-AF, 0.0070 mM for MRT-AP, 0.0089 mM for MRT-AT, and 0.0043 mM for MRT-AR Moloney murine leukemia virus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
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single band with a molecular mass of 75 kDa for Moloney murine leukemia virus reverse transcriptase and the mutants MRT-D224A and MRT-D524A, SDS-PAGE. Major single bands with molecular masses in the range of 65-73 kDa, each corresponding to their calculated molecular masses (70203, 71342, 71973 and 66387 Da), respectively, for the Moloney murine leukemia virus/avian myeloblastosis virus chimeric reverse transcriptases (MRT-AF, MRT-AP, MRT-AT, and MRT-AR) Moloney murine leukemia virus
additional information
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two bands with molecular masses of 95 kDa and 63 kDa for Avian myeloblastosis virus reverse transcriptase (purified from Avian myeloblastosis virus), SDS-PAGE. Major single bands with molecular masses in the range of 65-73 kDa, each corresponding to their calculated molecular masses (70203, 71342, 71973 and 66387 Da), respectively, for the Moloney murine leukaemia virus/Avian myeloblastosis virus chimeric reverse transcriptases (MRT-AF, MRT-AP, MRT-AT, and MRT-AR) Avian myeloblastosis virus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Avian myeloblastosis virus RNase H activity is determined by measuring the [3H]oligo(rA) released from [3H]poly(rA)*p(dT)45 ?
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?
additional information Moloney murine leukemia virus RNase H activity is determined by measuring the [3H]oligo(rA) released from [3H]poly(rA)*p(dT)45 ?
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?

Organism

Organism UniProt Comment Textmining
Avian myeloblastosis virus
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-
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Moloney murine leukemia virus P03355
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-

Purification (Commentary)

Purification (Comment) Organism
-
Avian myeloblastosis virus
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Moloney murine leukemia virus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
specific activity of Avian myeloblastosis virus reverse transcriptase at 37°C is 192000 U/mg RNase H, specific activity of the Moloney murine leukaemia virus/Avian myeloblastosis virus chimeric reverse transcriptases: MRT-AF = 10000 U/mg RNase H, MRT-AP U/mg = 10000 U/mg RNase H, MRT-AT U/mg = 13000 U/mg RNase H, and MRT-AR U/mg = 11000 U/mg RNase H. One unit (U) is defined as the amount which produces 1 pmol oligo(rA) (the amount is expressed as that of the total nucleotide) of 10%(w/v) trichloroacetic acid-soluble RNA in 20 min Avian myeloblastosis virus
additional information
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specific activity of Moloney murine leukemia virus reverse transcriptase at 37°C is 53000 U/mg RNase H, specific activity of the MRT-D224A mutant = 42000 U/mg RNase H, specific activity of the Moloney murine leukemia virus/avian myeloblastosis virus chimeric reverse transcriptases: MRT-AF = 10000 U/mg RNase H, MRT-AP U/mg = 10000 U/mg RNase H, MRT-AT U/mg = 13000 U/mg RNase H, and MRT-AR U/mg = 11000 U/mg RNase H. One unit (U) is defined as the amount which produces 1 pmol oligo(rA) (the amount is expressed as that of the total nucleotide) of 10%(w/v) TCA-soluble RNA in 20 min Moloney murine leukemia virus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information RNase H activity is determined by measuring the [3H]oligo(rA) released from [3H]poly(rA)*p(dT)45 Avian myeloblastosis virus ?
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?
additional information RNase H activity is determined by measuring the [3H]oligo(rA) released from [3H]poly(rA)*p(dT)45 Moloney murine leukemia virus ?
-
?

Synonyms

Synonyms Comment Organism
RNase H domain of reverse transcriptase Avian myeloblastosis virus
RNase H domain of reverse transcriptase Moloney murine leukemia virus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
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Avian myeloblastosis virus reverse transcriptase shows a curve with the optimal temperature range of around 44-50°C Avian myeloblastosis virus
44 50 Moloney murine leukemia virus reverse transcriptase shows a curve with the optimal temperature range of around 44-50°C Moloney murine leukemia virus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
additional information
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RNase H activities of Moloney murine leukemia virus reverse transcriptase and the chimeric reverse transcriptases at various temperatures ranging 30-60°C Moloney murine leukemia virus
44 50 RNase H activities of Avian myeloblastosis virus reverse transcriptase and the chimeric reverse transcriptases at various temperatures ranging 30-60°C Avian myeloblastosis virus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
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all chimeric reverse transcriptases are more thermally stable than Avian myeloblastosis virus reverse transcriptase in the RNase H reaction Avian myeloblastosis virus
additional information
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all chimeric reverse transcriptases are more thermally stable than Moloney murine leukemia virus reverse transcriptase in the RNase H reaction Moloney murine leukemia virus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information kcat value of Avian myeloblastosis virus reverse transcriptase (RNase H reaction) is above 0.025 s-1. The kcat values for different Moloney murine leukaemia virus/Avian myeloblastosis virus chimeric reverse transcriptases with poly(rA)*p(dT)45 as a substrate: 6.3 1/s for MRT-AF, 3.8 1/s for MRT-AP, 4.6 1/s for MRT-AT, and 2.4 1/s for MRT-AR Avian myeloblastosis virus
additional information
-
additional information kcat value of Moloney murine leukemia virus reverse transcriptase (RNase H reaction) above 3 ms-1. The kcat values for different Moloney murine leukemia virus/avian myeloblastosis virus chimeric reverse transcriptases with poly(rA)*p(dT)45 as a substrate: 6.3 1/s for MRT-AF, 3.8 1/s for MRT-AP, 4.6 1/s for MRT-AT, and 2.4 1/s for MRT-AR Moloney murine leukemia virus