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Literature summary extracted from

  • Jackman, J.E.; Phizicky, E.M.
    Identification of critical residues for G-1 addition and substrate recognition by tRNA(His) guanylyltransferase (2008), Biochemistry, 47, 4817-4825.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.79 expression in Escherichia coli Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.79 D131A 2–3% of wild-type activity Saccharomyces cerevisiae
2.7.7.79 D153A poorly expressed. D153A is stable, but its expression is somehow toxic to Escherichia coli. 312% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 D47A 54.7% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 D68A 59% of wild-type G(-1) addition activity. The mutation causes a dramatic decrease in the rigorous specificity of Thg1 for tRNAHis. This single alteration enhances the kcat/KM for ppp-tRNAPhe by nearly 100-fold relative to that of wild-type Thg1. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 D77A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 E13A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 E192A 2–3% of wild-type activity Saccharomyces cerevisiae
2.7.7.79 E75A 105% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 E78A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 G74A 44.4% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 H155A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 K190A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 K211A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 K44A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 K96A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 N157A 2–3% of wild-type activity Saccharomyces cerevisiae
2.7.7.79 N161A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 N201A less than 0.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 N46A 22.6% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 P45A 62.3% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 R133A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 R150A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 R27A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 R93A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 W113A/I156V 40.1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 Y146A 2–3% of wild-type activity Saccharomyces cerevisiae
2.7.7.79 Y160A 0.1%-1% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
2.7.7.79 Y202A poorly expressed, The Y202A variant is present at very low levels in soluble crude extracts and yields nearly undetectable levels of protein after purification, suggesting that this alteration leads to lack of overall stability Saccharomyces cerevisiae
2.7.7.79 Y40A 42.9% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.79 0.00016
-
 ppp-tRNAHis pH 7.5, 22°C, wild-type enzyme Saccharomyces cerevisiae
2.7.7.79 0.00024
-
 ppp-tRNAHis pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae
2.7.7.79 0.00041
-
 ppp-tRNAPhe pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae
2.7.7.79 0.0013
-
 ppp-tRNALeu pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae
2.7.7.79 0.025
-
 ppp-tRNAPhe pH 7.5, 22°C, wild-type enzyme Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.79 p-tRNAHis + ATP + GTP Saccharomyces cerevisiae
-
 pppGp-tRNAHis + AMP + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.79 Saccharomyces cerevisiae P53215
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.79 recombinant enzyme Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.79 p-tRNAHis + ATP + GTP
-
 Saccharomyces cerevisiae pppGp-tRNAHis + AMP + diphosphate
-
 ?
2.7.7.79 ppp-tRNAHis + GTP
-
 Saccharomyces cerevisiae pppGp-tRNAHis + diphosphate
-
 ?
2.7.7.79 ppp-tRNALeu + GTP the D68A mutation causes a dramatic decrease in the rigorous specificity of Thg1 for tRNAHis. This single alteration enhances the kcat/KM for ppp-tRNALeu by 38-fold relative to that of wild-type Thg1 Saccharomyces cerevisiae ?
-
 ?
2.7.7.79 ppp-tRNAPhe + GTP the D68A mutation causes a dramatic decrease in the rigorous specificity of Thg1 for tRNAHis. This single alteration enhances the kcat/KM for ppp-tRNAPhe by nearly 100-fold relative to that of wild-type Thg1 Saccharomyces cerevisiae ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.79 Thg1p
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.79 22
-
 assay at room temperature Saccharomyces cerevisiae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.79 0.00005
-
 ppp-tRNALeu pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae
2.7.7.79 0.00012
-
 ppp-tRNAPhe pH 7.5, 22°C, wild-type enzyme Saccharomyces cerevisiae
2.7.7.79 0.000172
-
 ppp-tRNAPhe pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae
2.7.7.79 0.00089
-
 ppp-tRNAHis pH 7.5, 22°C, wild-type enzyme Saccharomyces cerevisiae
2.7.7.79 0.0011
-
 ppp-tRNAHis pH 7.5, 22°C, mutant enzyme D68A Saccharomyces cerevisiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.79 7.5
-
 assay at Saccharomyces cerevisiae