Information on EC 6.1.1.27 - O-phospho-L-serine-tRNA ligase

Word Map on EC 6.1.1.27
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 enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.1.1.27
-
RECOMMENDED NAME
GeneOntology No.
O-phospho-L-serine-tRNA ligase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + O-phospho-L-serine + tRNACys = AMP + diphosphate + O-phospho-L-seryl-tRNACys
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
-
L-cysteine biosynthesis II (tRNA-dependent)
-
-
serine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
O-phospho-L-serine:tRNACys ligase (AMP-forming)
In organisms like Archaeoglobus fulgidus lacking EC 6.1.1.16 (cysteine---tRNA ligase) for the direct Cys-tRNACys formation, Cys-tRNACys is produced by an indirect pathway, in which EC 6.1.1.27 (O-phosphoseryl-tRNA ligase) ligates O-phosphoserine to tRNACys, and EC 2.5.1.73 (O-phospho-L-seryl-tRNA: Cys-tRNA synthase) converts the produced O-phospho-L-seryl-tRNACys to Cys-tRNACys. The SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism [1]. Methanosarcina mazei can use both pathways, the direct route using EC 6.1.1.16 (cysteine---tRNA ligase) and the indirect pathway with EC 6.1.1.27 and EC 2.5.1.73 (O-phospho-L-seryl-tRNA: Cys-tRNA synthase) [2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
no activity in Methanobrevibacter smithii
-
-
-
Manually annotated by BRENDA team
no activity in Methanosphaera stadtmanae
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + O-phospho-L-serine + tRNAAmber
AMP + diphosphate + O-phospho-L-serine-tRNAAmber
show the reaction diagram
recognition of U34 and C35 of tRNAAmber by mutant E418N/E420N, no activity with wild-type SepRS, overview
-
-
?
ATP + O-phospho-L-serine + tRNAAmber
AMP + diphosphate + O-phospho-L-seryl-tRNAAmber
show the reaction diagram
-
mutant D418N/D420N/T423V
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
show the reaction diagram
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
show the reaction diagram
ATP + O-phospho-L-serine + tRNAOpal
AMP + diphosphate + O-phospho-L-serine-tRNAOpal
show the reaction diagram
recognition of U34 and C35 of tRNAOpal by mutant E418N/E420N, no activity with wild-type SepRS, overview
-
-
?
ATP + O-phospho-L-serine + tRNAOpal
AMP + diphosphate + O-phospho-L-seryl-tRNAOpal
show the reaction diagram
-
mutant D418N/D420N/T423V
-
-
?
ATP + O-phospho-L-threonine + tRNACys
AMP + diphosphate + O-phospho-L-threonyl-tRNACys
show the reaction diagram
additional information
?
-
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
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
show the reaction diagram
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00097
m1G37-tRNACys
-
60°C, pH 6.0, steady-state kinetics
-
0.04 - 0.27
O-phospho-L-serine
0.93 - 8.4
O-phospho-L-threonine
0.0011 - 0.269
tRNACys
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24
m1G37-tRNACys
Methanocaldococcus jannaschii
-
60°C, pH 6.0, steady-state kinetics
-
0.45 - 11.2
O-phospho-L-serine
0.0054 - 0.021
O-phospho-L-threonine
0.008 - 1
tRNACys
additional information
additional information
Methanosarcina mazei
-
turnover number for tRNACys isoacceptor
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
recombinant enzyme, ATP-diphosphate exchange activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60909
-
4 * 60909, calculated from sequence; 4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry
68992
-
4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry
250000
-
gel filtration; recombinant enzyme,native PAGE, and gel filtration
255400
-
recombinant enzyme, mass spectrometry
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
SepRS tetramer in complex with tRNACys and O-phosphoserine, selenomethionine SAD method, and SepRS-tRNACys binary complex, 0.001 ml of 6-8 mg/ml protein in 10 mM Tris-HCl buffer, pH 8.0, 5 mM MgCl2, 150 mM NaCl and 5 mM 2-mercaptoethanol, and 2 mM O-phospho-L-serine, mixed with 0.001 ml reservoir solution containing 8% w/v PEG 6000 and 1.2 M NaCl, 20°C, cryoprotection by 22% v/v glycerol, X-ray diffraction structure determination and analysis at 2.6 A and 2.8 A resolution, respectively, modeling, determination of crystal structures of SepRS(E418N/E420N)-tRNAOpal-O-phosphoserine and SepRS(E418N/E420N)-tRNAAmber-O-phosphoserine at 3.2 and 3.3 resolutions, respectively
tRNA-free SepRS, hanging drop vapor diffusion method, 0.001 ml of protein solution mixed with 0.001 ml reservoir solution containing 11.25% w/v PEG 4,000, 75 mM sodium citrate, 75 mM N-(2-acetamido)iminodiacetic acid-NaOH buffer, pH 6.7, versus 1 ml reservoir solution, 20°C, cryoprotection by 22% v/v glycerol, X-ray diffraction structure determination and analysis at 3.6 A resolution, modeling
-
hanging-drop vapor diffusion, 3.2 A resolution
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant
recombinant enzyme; recombinant His-tagged enzyme from Escherichia coli by anion exchange chromatography, gel filtration, and nickel affinity chromatography
-
recombinant; recombinant His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity by nickel affinity chromatography, the His-tag is cleaved off
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli; expression of His-tagged enzyme in Escherichia coli
-
expression of His-tagged wild-type and mutant enzymes in Escherichia coli
-
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), overexpression of SeMet-labeled SepRS in Escherichia coli strain B834(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D418N/D420N/T423V
-
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme, it is active with tRNA substrate containing unusual residues 7-(2-thienyl)-imidazo[4,5-b]pyridine and pyrrole-2-carbaldehyde in the anticodon, overview
E418D
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418D/E420D
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418D/E420Q
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418N
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418N/E420D
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418N/E420N
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418N/E420N/T423V
E418N/E420Q
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418Q
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418Q/E420D
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418Q/E420N
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418Q/E420Q
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E420D
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E420K
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E420N
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E420Q
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E420R
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
T307S
-
mutant reveals a 3.2fold improvement in kcat/Km for phosphothreonyl adenylate synthesis, as compared with wild-type SepRS. The mutant is unable to transfer phosphothreonine to tRNACys at greater than 10% plateau levels; site-directed mutagenesis, the mutant shows increased activity with phosphothreonine, thus reduced substrate specificity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
the mutant SepRS-tRNA pairs may be useful for translational incorporation of O-phosphoserine into proteins in response to the stop codons UGA and UAG, so that it could ligate O-phosphoserine to a suppressor tRNA for genetic-code expansion
Show AA Sequence (138 entries)
Please use the Sequence Search for a specific query.