Information on EC 2.1.1.220 - tRNA (adenine58-N1)-methyltransferase

Word Map on EC 2.1.1.220
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
2.1.1.220
-
RECOMMENDED NAME
GeneOntology No.
tRNA (adenine58-N1)-methyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + adenine58 in tRNA = S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
tRNA methylation (yeast)
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:tRNA (adenine58-N1)-methyltransferase
The enzyme specifically methylates adenine58 in tRNA. The methylation of A58 is critical for maintaining the stability of initiator tRNAMet in yeast [3].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + adenine58 in human tRNA3Lys
S-adenosyl-L-homocysteine + N1-methyladenine58 in human tRNA3Lys
show the reaction diagram
Q96FX7 and Q9UJA5
-
-
-
?
S-adenosyl-L-methionine + adenine58 in initiator tRNAMet
S-adenosyl-L-homocysteine + N1-methyladenine58 in initiator tRNAMet
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in tRNA
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in tRNA3Lys
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA3Lys
show the reaction diagram
-
-
-
?
S-adenosyl-L-methionine + adenine58 in tRNAGGUThr
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in tRNAPhe
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNAPhe
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in yeast initiator tRNAMet
S-adenosyl-L-homocysteine + N1-methyladenine58 in yeast initiator tRNAMet
show the reaction diagram
Q96FX7 and Q9UJA5
no methylation of the A58U mutant of yeast initiator tRNA(Met)
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + adenine58 in initiator tRNAMet
S-adenosyl-L-homocysteine + N1-methyladenine58 in initiator tRNAMet
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in tRNA
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA
show the reaction diagram
S-adenosyl-L-methionine + adenine58 in tRNA3Lys
S-adenosyl-L-homocysteine + N1-methyladenine58 in tRNA3Lys
show the reaction diagram
Q96FX7
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Mg2+
-
1-10 mM, decrease in methylation rate. 95% inhibition at 10 mM
additional information
-
methylation at A58 is not affected by the presence of ribothymidine at position 54
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000025
adenine58 in initiator tRNAMet
-
pH 8.0, 30°C, purified FlagGcd10pyGcd14p complex
-
0.00013
adenine58 in tRNAPhe
-
pH 7.5-8.0, 55°C, wild-type Thermus thermophilus tRNAPhe as substrate
-
0.005
S-adenosyl-L-methionine
-
pH 8.0, 30°C, purified FlagGcd10pyGcd14p complex
additional information
additional information
-
kinetics for diverse tRNAPhe variants, overview
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8
-
assay at
7.6
Q96FX7 and Q9UJA5
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
from adrenal gland
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28582
-
4 * 28582, calculation from sequence, composed of two types of subunits (Gcd14p and Gcd10p)
28700
x * 29000, recombinant enzyme, SDS-PAGE, x * 28700, about, sequence calculation
29000
x * 29000, recombinant enzyme, SDS-PAGE, x * 28700, about, sequence calculation
31000
-
4 * 31000, SDS-PAGE
115000
-
gel filtration
150000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 29000, recombinant enzyme, SDS-PAGE, x * 28700, about, sequence calculation
heterotetramer
-
dimers of tightly assembled heterodimers, formed by TrmI-6 and TrmI-61 subunits, interactions and structure analysis, overview
homotetramer
-
4 * 31000, SDS-PAGE
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme in complex with S-adenosyl-L-methionine, sitting drop vapor diffusion method, mixing of 0.001 ml of 10-12 mg/ml protein in 20 mM Tris-HCl buffer, pH 8.0, containing 150 mM NaCl, 1 mM DTT, and 2 mM S-adenosyl-L-methionine, with 0.001 ml of reservoir solution containing 0.1 M Tris-HCl buffer, pH 8.4, and 20% ethanol, at 20°C, cryoprotection in 0.1 M Tris-HCl buffer, pH 8.4, 20% ethanol, and 35% ethylene glycol, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement method, using the coordinates of TrmI from Thermotoga maritima, PDB ID 1O54, as the starting model
TrmI complexed with S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
human enzyme tRNA m1A58 MTase in complex with human tRNA3Lys and cofactors S-adenosyl-L-methionine or S-adenosyl-L-homocysteine, hanging drop vapor diffusion method, mixing of 0.001 ml of 4.8 mg/ml protein in 50 mM HEPES, pH 7.5, 0.0664 mM tRNA3Lys, 2 mM S-adenosyl-L-methionine or S-adenosyl-L-homocysteine, and 1 mM MgCl2, with 0.001 ml of reservoir solution containing 0.1 M Na acetate, pH 4.8-5.0, 2% w/v PEG 4000 and 15% v/v methyl-2,4-pentanediol, 16°C, 4-7 days, X-ray diffraction structure determination and analysis at 2.2-4.0 A resolution, molecular replacement
TrmI-61 protein complexed with S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 2.5 A resolution
-
crystal structure of Rv2118c in complex with S-adenosyl-L-methionine has been determined at 1.98 A resolution
TrmI complexed with S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 1.98 A resolution
-
TrmI protein complexed as tetramer with S-adenosyl-L-homocysteine or as monomer with S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 2.05-2.6 A or 1.6 A resolution, respectively
-
purified recombinant His-tagged TRM6-TRM61 complex, from 0.1 M KCl, 0.1 M Tris-HCl, pH 8.0, 25% w/v PEG 2000 MME, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement
-
ligand-free TrmI, X-ray diffraction structure determination and analysis at 1.65 A resolution
-
purified enzyme mutant D170A and Y78A in complex with S-adenosyl-L-methionine, hanging drop vapor diffusion method, mixing of 10 mg/ml protein in 20 mM Tris-HCl buffer, pH 8.0, 100 mM KCl, and 2mM S-adenosyl-L-methionine with reservoir solution containing 2.4 M ammonium sulfate and 10% v/v isopropanol for mutant D170A and 2.1 M ammonium sulfate and 8% v/v isopropanol for mutant Y78A, X-ray diffraction structure determination and analysis at 3.1 A and 2.6 A resolution, respectively. Crystallization assays of enzyme TrmI Y194A lead to poorly diffracting crystals
-
sitting-drop vapor-diffusion method at 19°C. Crystal structure of TrmI, in complex with S-adenosyl-L-homocysteine, is determined at 1.7 A resolution. The conserved residues that form the catalytic cavity (D170, Y78, and Y194) are essential for fashioning an optimized shape of the catalytic pocket
-
TrmI protein complexed with S-adenosyl-L-homocysteine, X-ray diffraction structure determination and analysis at 1.7 A resolution
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
coexpression of catalytic subunit Trm61p and non-catalytic subunit Trm6p in Saccharomyces cerevisiae, purification of the hTrm6p/hTrm61p complexes
Q96FX7 and Q9UJA5
recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the tag by 3C protease, and gel filtration
recombinant enzyme from Escherichia coli strain Rosetta (DE3) by heat treatment at 70°C for 30 min, anion exchange and hydrophobic interaction chromatography, and dialysis, followed by cation exchange chromatography, gel filtration, and ultrafiltration
recombinant His-tagged TRM6 and TRM61 from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
coexpression of catalytic subunit Trm61p and non-catalytic subunit Trm6p in Saccharomyces cerevisiae
Q96FX7 and Q9UJA5
expression in Escherichia coli
gene aq_311, recombinant expression in Escherichia coli strain Rosetta (DE3)
genes GCD10 and GCD14 encoding TRM6 and TRM61, expression of His-tagged proteins in Escherichia coli strain Rosetta (DE3)
-
recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D170A
-
site-directed mutagenesis, mutation of a conserved active site residue
Y194A
-
site-directed mutagenesis, crystallization assays of TrmI Y194A lead to poorly diffracting crystals
Y78A
-
site-directed mutagenesis, mutation of a conserved active site residue. The structure of TrmI Y78A catalytic domain is unmodified regarding the binding of the SAM co-factor and the conformation of residues potentially interacting with the substrate adenine, as compared to the wild-type structure. The structure of the D170A mutant shows a flexible active site with one loop occupying in part the place of the co-factor and the second loop moving at the entrance to the active site
D170A
-
site-directed mutagenesis, mutation of a conserved active site residue
-
Y194A
-
site-directed mutagenesis, crystallization assays of TrmI Y194A lead to poorly diffracting crystals
-
Y78A
-
site-directed mutagenesis, mutation of a conserved active site residue. The structure of TrmI Y78A catalytic domain is unmodified regarding the binding of the SAM co-factor and the conformation of residues potentially interacting with the substrate adenine, as compared to the wild-type structure. The structure of the D170A mutant shows a flexible active site with one loop occupying in part the place of the co-factor and the second loop moving at the entrance to the active site
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
the enzyme crystal structures serve as templates for design of inhibitors that can be used to test tRNA m1A58 MTase's impact on retroviral priming and transcription
Show AA Sequence (237 entries)
Please use the Sequence Search for a specific query.