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Literature summary extracted from

  • Klarmann, G.J.; Eisenhauer, B.M.; Zhang, Y.; Gotte, M.; Pata, J.D.; Chatterjee, D.K.; Hecht, S.M.; Le Grice, S.F.
    Investigating the "steric gate" of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase by targeted insertion of unnatural amino acids (2007), Biochemistry, 46, 2118-2126.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.49 variants of p66 subunit of reverse transcriptase containing meta-Tyr, nor-Tyr, aminomethyl-Phe, and 1-naphthyl-Tyr and 2-naphthyl-Tyr are produced in an Escherichia coli coupled transcription/translation system. Mutant p66 subunits are reconstituted with wild-type p51 subunit of reverse transcriptase Human immunodeficiency virus 1

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.49 Y1152-naphthyl-Tyr mutant of the p66 subunit of reverse transcriptase reconstituted with wild-type p51 subunit. Mutant enzyme inefficiently incorporates dCTP at low concentrations and is kinetically slower with all dCTP analogues tested. 5fold less efficient for dCTP incorporation and 15fold less efficient for 2',3'-dideoxy-CTP incorporation compared to the wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 Y115aminomethyl-Phe mutant of the p66 subunit of reverse transcriptase reconstituted with wild-type p51 subunit. Mutant enzyme incorporates dCTP more efficiently than the wild-type and is resistant to the chain terminator (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate when examined in a steady-state fidelity assay. Mutant enzyme incorporates very little 3TCMP, even when the concentration of the chain terminator was 100times that of the dCTP Human immunodeficiency virus 1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.49 0.0014
-
 dCTP wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.00217
-
 2',3'-dideoxy-CTP wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.00273
-
 dCTP wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.003
-
 2',3'-dideoxy-CTP mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 0.00358
-
 2',3'-dideoxy-CTP mutant enzyme Y1152-naphthyl-Tyr Human immunodeficiency virus 1
2.7.7.49 0.00682
-
 dCTP mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 0.01123
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.0156
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate mutant enzyme Y1152-naphthyl-Tyr Human immunodeficiency virus 1
2.7.7.49 0.0275
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.49 Human immunodeficiency virus 1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.49 variants of p66 subunit of reverse transcriptase containing meta-Tyr, nor-Tyr, aminomethyl-Phe, and 1-naphthyl-Tyr and 2-naphthyl-Tyr are produced in an Escherichia coli coupled transcription/translation system. Mutant p66 subunits are reconstituted with wild-type p51 subunit of reverse transcriptase and purified Human immunodeficiency virus 1

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.49 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate + 2-naphthyl-Tyr115 of p66 reverse transcriptase
-
 Human immunodeficiency virus 1 ?
-
 ?
2.7.7.49 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate + aminomethyl-Phe115 of p66 reverse transcriptase
-
 Human immunodeficiency virus 1 ?
-
 ?
2.7.7.49 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate + DNAn
-
 Human immunodeficiency virus 1 diphosphate + ?
-
 ?
2.7.7.49 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate + Tyr115 of p66 reverse transcriptase
-
 Human immunodeficiency virus 1 ?
-
 ?
2.7.7.49 2',3'-dideoxy-CTP + DNAn
-
 Human immunodeficiency virus 1 diphosphate + DNAn+1
-
 ?
2.7.7.49 dCTP + DNAn
-
 Human immunodeficiency virus 1 diphosphate + DNAn+1
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.49 p66 RT
-
 Human immunodeficiency virus 1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.49 0.042
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate mutant enzyme Y1152-naphthyl-Tyr Human immunodeficiency virus 1
2.7.7.49 0.042
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphat mutant enzyme Y1152-naphthyl-Tyr Human immunodeficiency virus 1
2.7.7.49 0.077
-
 2',3'-dideoxy-CTP mutant enzyme Y1152-naphthyl-Tyr Human immunodeficiency virus 1
2.7.7.49 0.15
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.15
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphat wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.16
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphate mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 0.16
-
 (-)-beta-2',3'-dideoxy-3'-thiacytidine triphosphat mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 0.49
-
 2',3'-dideoxy-CTP mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 0.67
-
 2',3'-dideoxy-CTP wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 0.76
-
 dCTP mutant enzyme Y115aminomethyl-Phe Human immunodeficiency virus 1
2.7.7.49 1.25
-
 dCTP wild-type enzyme Human immunodeficiency virus 1
2.7.7.49 1.64
-
 dCTP wild-type enzyme Human immunodeficiency virus 1