Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.7.79 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
Search for more literature for 2.7.7.79

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
D131A 2–3% of wild-type activity Saccharomyces cerevisiae
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
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
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
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
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
E192A 2–3% of wild-type activity Saccharomyces cerevisiae
E75A 105% of wild-type G(-1) addition activity. G(-) i.e. the extra nucleotide posttranscriptionally added in position (-1) of tRNA Saccharomyces cerevisiae
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
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
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
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
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
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
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
N157A 2–3% of wild-type activity Saccharomyces cerevisiae
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
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
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
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
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
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
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
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
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
Y146A 2–3% of wild-type activity Saccharomyces cerevisiae
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
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
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P53215
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
p-tRNAHis + ATP + GTP
-
 Saccharomyces cerevisiae pppGp-tRNAHis + AMP + diphosphate
-
 ?
ppp-tRNAHis + GTP
-
 Saccharomyces cerevisiae pppGp-tRNAHis + diphosphate
-
 ?
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 ?
-
 ?
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

Synonyms Comment Organism
Thg1p
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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