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Literature summary for 2.4.99.17 extracted from

  • Van Lanen, S.G.; Kinzie, S.D.; Matthieu, S.; Link, T.; Culp, J.; Iwata-Reuyl, D.
    tRNA modification by S-adenosylmethionine:tRNA ribosyltransferase-isomerase. Assay development and characterization of the recombinant enzyme (2003), J. Biol. Chem., 278, 10491-10499.
    View publication on PubMed
Search for more literature for 2.4.99.17

Activating Compound

Activating Compound Comment Organism Structure
Carbonate 100% activity in the presence of 25 mM carbonate Escherichia coli
DNA activity is highest in the presence of 0.2 mg/ml DNA Escherichia coli
EDTA 100% activity in the presence of 100 mM EDTA Escherichia coli
Sodium acetate 100% activity in the presence of 750 mM sodium acetate Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH5apha cells Escherichia coli

General Stability

General Stability Organism
replacing Tris buffer with glycyclglycine increases activity 2fold Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
CaCl2 43% residual activity at 10 mM Escherichia coli
KCl 28% residual activity at 100 mM Escherichia coli
Mg2+ Mg2+ in millimolar concentrations inhibits enzyme activity but has little or no effect in the submillimolar range Escherichia coli
Mn2+ Mn2+ in millimolar concentrations inhibits enzyme activity but has little or no effect in the submillimolar range Escherichia coli
Na2PO4 80% residual activity at 25 mM Escherichia coli
Na2SO4 33% residual activity at 25 mM Escherichia coli
NaCl 25% residual activity at 100 mM Escherichia coli
NaSCN 60% residual activity at 0.01 mM Escherichia coli
potassium fluoride 78% residual activity at 500 mM Escherichia coli
Tris-HCl 11% residual activity at 100 mM Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0015
-
 7-aminomethyl-7-deazaguanine in tRNA(Tyr) in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli
0.0377
-
 7-aminomethyl-7-deazaguanine in minihelix RNA in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli
0.1014
-
 S-adenosyl-L-methionine in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + 7-aminomethyl-7-deazaguanine in tRNA(Tyr) Escherichia coli
-
 L-methionine + adenine + epoxyqueuosine in tRNA(Tyr)
-
 ir

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
glutathione-Sepharose column chromatography and POROS HQ column chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + 7-aminomethyl-7-deazaguanine in minihelix RNA efficient substrate Escherichia coli L-methionine + adenine + epoxyqueuosine in minihelix RNA
-
 ir
S-adenosyl-L-methionine + 7-aminomethyl-7-deazaguanine in tRNA(Tyr)
-
 Escherichia coli L-methionine + adenine + epoxyqueuosine in tRNA(Tyr)
-
 ir

Synonyms

Synonyms Comment Organism
QueA
-
 Escherichia coli
S-adenosylmethionine:tRNA ribosyltransferase-isomerase
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.042
-
 S-adenosyl-L-methionine in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli
0.042
-
 7-aminomethyl-7-deazaguanine in tRNA(Tyr) in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli
0.245
-
 7-aminomethyl-7-deazaguanine in minihelix RNA in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.7
-
 optimal activity at pH 8.7 in buffers containing various oxyanions, including acetate, carbonate, EDTA, and phosphate Escherichia coli

General Information

General Information Comment Organism
metabolism the enzyme catalyzes the penultimate step in the de novo biosynthesis of queuosine Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.67
-
 7-aminomethyl-7-deazaguanine in minihelix RNA in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli
28.3
-
 7-aminomethyl-7-deazaguanine in tRNA(Tyr) in 100 mM glycylglycine (pH 8.7), 100 mM EDTA, 100 mM KCl, 0.5 mM dithiothreitol, at 37°C Escherichia coli