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Literature summary extracted from

  • Reiter, N.J.; Osterman, A.K.; Mondragon, A.
    The bacterial ribonuclease P holoenzyme requires specific, conserved residues for efficient catalysis and substrate positioning (2012), Nucleic Acids Res., 40, 10384-10393.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.26.5 genes rnpA and rnpB, recombinant expression of GST-tagged wild-type and mutant proteins and RNA in Escherichia coli strain BL21(DE3)pLysS Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
3.1.26.5 F17A site-directed mutagenesis of the protein part, the mutant shows 95% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 F17A/U52C site-directed mutagenesis of the protein part (F17A) and the RNA part (U52C), the mutant shows over 99% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 F21A site-directed mutagenesis of the protein part, the mutant shows 80% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 K51A site-directed mutagenesis of the protein part, the mutant shows 60% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 K53A site-directed mutagenesis of the protein part, the mutant shows 80% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 K56A site-directed mutagenesis of the protein part, the mutant shows 10% increased activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 K62A site-directed mutagenesis of the protein part, the mutant shows unaltered activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 K90A site-directed mutagenesis of the protein part, the mutant shows 70% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 additional information wild-type and mutant enzyme structure-function analysis, overview. Point mutations in the conserved protein loop (residues 52-57) have either no or modest effects on catalytic efficiency. Similarly, amino acid changes in the RNR region, which represent the most conserved region of bacterial RNase P proteins, exhibit negligible changes in catalytic efficiency Thermotoga maritima
3.1.26.5 R14A site-directed mutagenesis of the protein part, the mutant shows 60% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R15A site-directed mutagenesis of the protein part, the mutant shows 70% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R52A site-directed mutagenesis of the protein part, the mutant shows 60% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R59A site-directed mutagenesis of the protein part, the mutant shows 20% increased activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R60A site-directed mutagenesis of the protein part, the mutant shows 30% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R65A site-directed mutagenesis of the protein part, the mutant shows 50% increased activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R89A site-directed mutagenesis of the protein part, the mutant shows 94% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 R89A/U52C site-directed mutagenesis of the protein part (R89A) and the RNA part (U52C), the mutant shows over 99% reduced activity compared to the wild-type enzyme Thermotoga maritima
3.1.26.5 U52C site-directed mutagenesis of the RNA part, the mutant shows 92% reduced activity compared to the wild-type enzyme Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.26.5 additional information
-
 additional information Michaelis-Menten single-turnover reaction kinetics of wild-type and mutant enzymes, overview Thermotoga maritima

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.26.5 Mg2+ required, the active site includes at least two metal ions, RNA U52 nucleotide binds a metal ion at the active site Thermotoga maritima

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.26.5 additional information Thermotoga maritima the enzyme is an RNA-based enzyme primarily catalyzing 5'-end pre-tRNA processing ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.26.5 Thermotoga maritima Q9X1H4
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.1.26.5 ribonucleoprotein
-
 Thermotoga maritima

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.26.5 recombinant GST-tagged wild-type and mutant proteins and RNA from Escherichia coli strain BL21(DE3)pLysS Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.26.5 additional information the enzyme is an RNA-based enzyme primarily catalyzing 5'-end pre-tRNA processing Thermotoga maritima ?
-
 ?
3.1.26.5 pre-tRNAPhe + H2O
-
 Thermotoga maritima ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.26.5 More wild-type and mutant enzyme structure-function analysis, overview Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
3.1.26.5 RNase P
-
 Thermotoga maritima

General Information

EC Number General Information Comment Organism
3.1.26.5 additional information wild-type and mutant enzyme structure-function analysis, overview. RNA U52 and two bacterially conserved protein residues, F17 and R89, are essential for efficient Thermotoga maritima enzyme activity. The U52 nucleotide binds a metal ion at the active site, whereas F17 and R89 are positioned over 20 A from the cleavage site, probably making contacts with N-4 and N-5 nucleotides of the pretRNA 5'-leader Thermotoga maritima
3.1.26.5 physiological function the enzyme is an RNA-based enzyme primarily responsible for 5'-end pre-tRNA processing Thermotoga maritima

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.26.5 0.83
-
 pre-tRNAPhe recombinant mutant R89A/U52C, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 1
-
 pre-tRNAPhe recombinant mutant F17A/U52C, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 8.3
-
 pre-tRNAPhe recombinant mutant F17A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 10
-
 pre-tRNAPhe recombinant mutant R89A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 13
-
 pre-tRNAPhe recombinant mutant U52C, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 28.3
-
 pre-tRNAPhe recombinant mutant F21A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 40
-
 pre-tRNAPhe recombinant mutant K53A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 44.2
-
 pre-tRNAPhe recombinant mutant K90A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 54.2
-
 pre-tRNAPhe recombinant mutant R15A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 65
-
 pre-tRNAPhe recombinant mutant R14A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 66.7
-
 pre-tRNAPhe recombinant mutant K52A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 70
-
 pre-tRNAPhe recombinant mutant K51A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 120
-
 pre-tRNAPhe recombinant mutant R60A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 170
-
 pre-tRNAPhe recombinant mutant K62A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 180
-
 pre-tRNAPhe recombinant wild-type enzyme, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 181.7
-
 pre-tRNAPhe recombinant mutant K56A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 198.3
-
 pre-tRNAPhe recombinant mutant R59A, pH 7.5, 37°C Thermotoga maritima
3.1.26.5 261.7
-
 pre-tRNAPhe recombinant mutant R65A, pH 7.5, 37°C Thermotoga maritima