Any feedback?
Information on EC 6.1.1.27 - O-phospho-L-serine-tRNA ligase
for references in articles please use BRENDA:EC6.1.1.27Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
6.1.1.27 O-phospho-L-serine-tRNA ligaseIUBMB Comments
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 . 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) .
Specify your search results
Word Map
- 6.1.1.27
-
cys-trnacys
-
methanogen
- aminoacylation
- archaea
- sepcyss
-
cysteinyl-trna
-
sep-trna:cys-trna
- o-phosphoserine
-
anticodons
- aminoacyl-trnas
- maripaludis
-
trna-dependent
-
cysteinyl-trnacys
- selenocysteine
-
methanocaldococcus
- jannaschii
-
phosphoserylation
- pylrs
-
aarss
-
faithful
- mazei
-
methanosarcina
- gca
-
alpha4
-
web-based
-
biologists
- phenylalanyl-trna
- asnrs
- ilers
- biotechnology
The expected taxonomic range for this enzyme is: Archaea, Bacteria
Reaction Schemes
Synonyms
seprs, o-phosphoseryl-trna synthetase, phosphoseryl-trna synthetase, phosphoseryl-trna synthetases, 
more
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
CysRS
O-phosphoseryl-tRNA synthetase
phosphoseryl-tRNA synthetase
SepRS
sepS
additional information
additional information
-
the enzyme is a class II tRNA synthetase and belongs to the PLP-dependent superfamily of enzymes
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + O-phospho-L-serine + tRNACys = AMP + diphosphate + O-phospho-L-seryl-tRNACys
anticodon recognition mechanism
ATP + O-phospho-L-serine + tRNACys = AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PATHWAY SOURCE
PATHWAYS
BRENDA
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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].
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + O-phospho-L-serine + m1G37-tRNACys
AMP + diphosphate + O-phospho-L-seryl-m1G37-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNAAmber
AMP + diphosphate + O-phospho-L-serine-tRNAAmber
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
-
mutant D418N/D420N/T423V
-
-
?
ATP + O-phospho-L-serine + tRNAOpal
AMP + diphosphate + O-phospho-L-serine-tRNAOpal
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
-
mutant D418N/D420N/T423V
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
tRNA substrate from Escherichia coli, wheat germ and Saccharomyces cerevisiae in a mixture, the catalytic domain of SepRS recognizes the negatively charged side chain of O-phosphoserine at a noncanonical site, using the dipole moment of a conserved alpha-helix, the unique C-terminal domain specifically recognizes the anticodon GCA of tRNACys, overview
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
phosphoseryl-tRNA synthetase is a natural non-standard aminoacyl-tRNA synthetase, which charges a non-standard amino acid, phosphoserine, to tRNACys containing a GCA anticodon for tRNA-dependent cysteine biosynthesis in some archaea
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
cognate substrate is tRNACys with the GCA anticodon, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryltRNA synthetase (SepRS) acylates tRNACys with phosphoserine (Sep), and Sep-tRNACys-tRNA synthase (SepCysS) converts the tRNA-bound phosphoserine to cysteine
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
SepRS differs from CysRS (EC 6.1.1.16) by recruiting the m1G37 modification as a determinant for aminoacylation, and in showing limited discrimination against mutations of conserved nucleotides. O-PhosphoseryltRNA synthetase and Sep-tRNACys-tRNA synthase bind the reaction intermediate O-phospho-L-serine-tRNACys tightly, and these two enzymes form a stable binary complex that promotes conversion of the intermediate to the product and sequesters the intermediate from binding to elongation factor EF-1a or infiltrating into the ribosome
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
SepRS differs from CysRS by recruiting the m1G37 modification as a determinant for aminoacylation, and in showing limited discrimination against mutations of conserved nucleotides. The enzyme requires the S-adenosylmethione-dependent formation of m1G37 in the anticodon loop for efficient aminoacylation
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
Methanococcus maripaludis encodes both the direct and indirect paths for Cys-tRNACys synthesis. SepS (encoding SepRS) can be deleted when the organism is grown in the presence of Cys
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
the enzyme charges tRNACys with a 3'-deoxyadenosine, but not tRNACys without A76 or with a 2'-deoxyadensosine
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
half-of-the-sites activity: the tetrameric enzyme binds two tRNAs. Only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
some methanogenic archaea synthesize Cys-tRNACys needed for protein synthesis using both a canonical cysteinyl-tRNA synthetase as well as a set of two enzymes that operate via a separate indirect pathway. In the indirect route, Sep-tRNACys is first synthesized by SepRS, and this misacylated intermediate is then converted to Cys-tRNACys by Sep-tRNA:Cys-tRNA synthase via a pyridoxal phosphate-dependent mechanism, structural basis for the tRNACys isoacceptor preferences of SepRS and CysRS, detailed overview
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
efficient phosphoserylation by SepRS requires methylation of tRNACys at the N1 position of G37 in the anticodon loop. Comparative aminoacylation kinetics by CysRS (EC 6.1.1.16) and SepRS reveals that each enzyme prefers a distinct tRNACys isoacceptor or pair of isoacceptors
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
half-of-the-sites activity: the tetrameric enzyme binds two tRNAs. Only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate. Efficient phosphoserylation by SepRS requires methylation of tRNACys at the N1 position of G37 in the anticodon loop
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
recognition determinants distinguishing the tRNAs reside in the globular core of the molecule. The enzyme also requires the S-adenosylmethione-dependent formation of m1G37 in the anticodon loop for efficient aminoacylation
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
the tetrameric enzyme binds two tRNAs and only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate. tRNACys binding to SepRS also enhances the capacity of the enzyme to discriminate among amino acids, indicating the existence of functional connectivity between the tRNA and amino acid binding sites of the enzyme
-
-
?
ATP + O-phospho-L-threonine + tRNACys
AMP + diphosphate + O-phospho-L-threonyl-tRNACys
-
low activity
-
-
?
ATP + O-phospho-L-threonine + tRNACys
AMP + diphosphate + O-phospho-L-threonyl-tRNACys
-
about 35% of the plateau aminoacylation observed with O-phospho-L-serine
-
-
?
additional information
?
-
two-step Cys-tRNACys formation: in organisms like Archaeoglobus fulgidus lacking a canonical cysteinyl-tRNA synthetase for the direct Cys-tRNACys formation, Cys-tRNACys is produced by the indirect pathway, in which the noncanonical O-phosphoseryl-tRNA synthetase, SepRS, ligates the noncanonical amino acid O-phosphoserine, Sep, to tRNACys, and the Sep-tRNA:Cys-tRNA synthase converts the produced Sep-tRNACys to Cys-tRNACys, overview, the SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism
-
-
?
additional information
?
-
-
substrate specificity, site-specific incorporation of phosphoserine into proteins by mutant D418N/D420N/T423V in response to the 7-(2-thienyl)-imidazo[4,5-b]pyridineUA or C7-(2-thienyl)-imidazo[4,5-b]pyridineA codons within mRNA, substrate binding structures and structural models for tRNA anticodon recognition, overview
-
-
?
additional information
?
-
two-step Cys-tRNACys formation: in organisms like Archaeoglobus fulgidus lacking a canonical cysteinyl-tRNA synthetase for the direct Cys-tRNACys formation, Cys-tRNACys is produced by the indirect pathway, in which the noncanonical O-phosphoseryl-tRNA synthetase, SepRS, ligates the noncanonical amino acid O-phosphoserine, Sep, to tRNACys, and the Sep-tRNA:Cys-tRNA synthase converts the produced Sep-tRNACys to Cys-tRNACys, overview, the SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism. RNA substrate specificity of wild-type and mutant enzymes, overview, structural insights into the first step of RNA-dependent cysteine biosynthesis, a two-step mechanism, in archaea
-
-
?
additional information
?
-
-
two-step Cys-tRNACys formation: in organisms like Methanococcus jannaschii lacking a canonical cysteinyl-tRNA synthetase for the direct Cys-tRNACys formation, Cys-tRNACys is produced by the indirect pathway, in which the noncanonical O-phosphoseryl-tRNA synthetase, SepRS, ligates the noncanonical amino acid O-phosphoserine, Sep, to tRNACys, and the Sep-tRNA:Cys-tRNA synthase converts the produced Sep-tRNACys to Cys-tRNACys, overview, the SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism
-
-
?
additional information
?
-
-
Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryl-tRNA synthetase acylates tRNACys with phosphoserine, and Sep-tRNA-Cys-tRNA synthase converts the tRNA-bound phosphoserine to cysteine. Methanocaldococcus jannaschii SepRS differs from CysRS by recruiting the m1G37 modification as a determinant for aminoacylation
-
-
?
additional information
?
-
-
kinetic and binding measurements show that both SepRS and Sep-tRNA-Cys-tRNA synthase, SepCysS, bind the reaction intermediate Sep-tRNACys tightly, and these two enzymes form a stable binary complex that promotes conversion of the intermediate to the product and sequesters the intermediate from binding to elongation factor EF-1alpha or infiltrating into the ribosome, mechanism of the binary complex, detailed overview
-
-
?
additional information
?
-
-
SepRS is able to discriminate against the noncognate amino acids glutamate, serine, and phosphothreonine without the need for a separate hydrolytic editing site
-
-
?
additional information
?
-
-
SepRS is able to discriminate against the noncognate amino acids glutamate, serine, and phosphothreonine without the need for a separate hydrolytic editing site. Determination of the ATP-diphosphate exchange activity. Serine and glutamate are poor substrates, overview
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-serine-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
phosphoseryl-tRNA synthetase is a natural non-standard aminoacyl-tRNA synthetase, which charges a non-standard amino acid, phosphoserine, to tRNACys containing a GCA anticodon for tRNA-dependent cysteine biosynthesis in some archaea
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryltRNA synthetase (SepRS) acylates tRNACys with phosphoserine (Sep), and Sep-tRNACys-tRNA synthase (SepCysS) converts the tRNA-bound phosphoserine to cysteine
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
Methanococcus maripaludis encodes both the direct and indirect paths for Cys-tRNACys synthesis. SepS (encoding SepRS) can be deleted when the organism is grown in the presence of Cys
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
the enzyme charges tRNACys with a 3'-deoxyadenosine, but not tRNACys without A76 or with a 2'-deoxyadensosine
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
-
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
half-of-the-sites activity: the tetrameric enzyme binds two tRNAs. Only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate
-
-
?
ATP + O-phospho-L-serine + tRNACys
AMP + diphosphate + O-phospho-L-seryl-tRNACys
-
some methanogenic archaea synthesize Cys-tRNACys needed for protein synthesis using both a canonical cysteinyl-tRNA synthetase as well as a set of two enzymes that operate via a separate indirect pathway. In the indirect route, Sep-tRNACys is first synthesized by SepRS, and this misacylated intermediate is then converted to Cys-tRNACys by Sep-tRNA:Cys-tRNA synthase via a pyridoxal phosphate-dependent mechanism, structural basis for the tRNACys isoacceptor preferences of SepRS and CysRS, detailed overview
-
-
?
additional information
?
-
two-step Cys-tRNACys formation: in organisms like Archaeoglobus fulgidus lacking a canonical cysteinyl-tRNA synthetase for the direct Cys-tRNACys formation, Cys-tRNACys is produced by the indirect pathway, in which the noncanonical O-phosphoseryl-tRNA synthetase, SepRS, ligates the noncanonical amino acid O-phosphoserine, Sep, to tRNACys, and the Sep-tRNA:Cys-tRNA synthase converts the produced Sep-tRNACys to Cys-tRNACys, overview, the SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism
-
-
?
additional information
?
-
-
two-step Cys-tRNACys formation: in organisms like Methanococcus jannaschii lacking a canonical cysteinyl-tRNA synthetase for the direct Cys-tRNACys formation, Cys-tRNACys is produced by the indirect pathway, in which the noncanonical O-phosphoseryl-tRNA synthetase, SepRS, ligates the noncanonical amino acid O-phosphoserine, Sep, to tRNACys, and the Sep-tRNA:Cys-tRNA synthase converts the produced Sep-tRNACys to Cys-tRNACys, overview, the SepRS/SepCysS pathway is the sole route for cysteine biosynthesis in the organism
-
-
?
additional information
?
-
-
Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryl-tRNA synthetase acylates tRNACys with phosphoserine, and Sep-tRNA-Cys-tRNA synthase converts the tRNA-bound phosphoserine to cysteine. Methanocaldococcus jannaschii SepRS differs from CysRS by recruiting the m1G37 modification as a determinant for aminoacylation
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.007
tRNACys
-
pH 7.5, 37°C, recombinant enzyme, native tRNACys
0.0097
tRNACys
-
pH 7.5, 37°C, recombinant enzyme, m1G37 tRNACys
0.151
tRNACys
-
50°C, pH 7.6, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon, mutant enzyme D418N D420N T423V
0.151
tRNACys
-
50°C, pH 7.6, tRNACys containing the C(pyrrole-2-carbaldehyde)A anticodon, mutant enzyme D418N D420N T423V
0.269
tRNACys
-
50°C, pH 7.6, tRNACys with the GCA anticodon, wild-type enzyme
additional information
additional information
-
kinetic analysis of Sep-tRNA formation, overview
-
additional information
additional information
-
Km-values for tRNACys isoacceptor
-
additional information
additional information
-
recombinant enzyme, ATP/diphosphate and aminoacylation kinetics, Michaelis-Menten kinetics
-
additional information
additional information
-
steady-state aminoacylation kinetics
-
additional information
additional information
-
steady-state and single-turnover kinetics, kinetic analysis, overview
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
turnover number for tRNACys isoacceptor
-
0.0471
tRNACys
-
50°C, pH 7.6, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon, mutant enzyme D418N D420N T423V
0.048
tRNACys
-
50°C, pH 7.6, tRNACys containing the C(pyrrole-2-carbaldehyde)A anticodon, mutant enzyme D418N D420N T423V
0.115
tRNACys
-
50°C, pH 7.6, tRNACys with the GCA anticodon, wild-type enzyme
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
recombinant enzyme, ATP-diphosphate exchange activity
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Show AA Sequence and Transmembrane Helices (559 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PDB
SCOP
CATH
UNIPROT
ORGANISM
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
250000
60909
68992
-
4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry
60909
-
4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
additional information
tetramer
-
SepRS contains two tRNACys molecules per tetramer indicating an asymmetry of the four identical subunits
tetramer
-
4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry
additional information
-
both SepRS and SepCysS are active as a monomer in theSepRSSepCysS binary complex
additional information
-
the tetrameric enzyme binds two tRNAs and only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate, active site titrations reveal calculation of 2.3 active sites per tetramer, overview
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
hanging drop vapor diffusion method, using 0.1 M Tris-HCl pH 7.0, 0.3M ammonium sulfate, 36% (w/v) 2-methyl-2,4-pentanediol
sitting drop vapor diffusion method, using 0.1 M MES pH 5.3, 2.37 M ammonium sulfate
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
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
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
additional information
E418N/E420N/T423V
site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme
E418N/E420N/T423V
-
efficiently charged phosphoserine to tRNA containing the (pyrrole-2-carbaldehyde)UA anticodon
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
T307S
-
site-directed mutagenesis, the mutant shows increased activity with phosphothreonine, thus reduced substrate specificity
additional information
engineering of SepRS to recognize tRNACys mutants with the anticodons UCA and CUA on the basis of the structure, phosphoserine ligation activity of the wild-type and mutant SepRSs for tRNACys, overview
additional information
-
mutation of the three anticodon nucleotides, G34, C35 and A36, as well as the next residue, G37, reduces the phosphoserylation activity
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography, HiTrap heparin column chromatography, and Superdex 200 gel filtration
recombinant
recombinant His-tagged enzyme from Escherichia coli by anion exchange chromatography, gel filtration, and nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity by nickel affinity chromatography, the His-tag is cleaved off
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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)
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Fukunaga, R.; Yokoyama, S.
Structural insights into the first step of RNA-dependent cysteine biosynthesis in archaea
Nat. Struct. Mol. Biol.
14
272-279
2007
Methanocaldococcus jannaschii, Archaeoglobus fulgidus (O30126)
brenda
Yuan, J.; Sheppard, K.; Soell, D.
Amino acid modifications on tRNA
Acta Biochim. Biophys. Sin. (Shanghai)
40
539-553
2008
Methanococcus maripaludis, no activity in Methanobrevibacter smithii, no activity in Methanosphaera stadtmanae
brenda
Hauenstein, S.I.; Perona, J.J.
Redundant synthesis of cysteinyl-tRNACys in Methanosarcina mazei
J. Biol. Chem.
283
22007-22017
2008
Methanosarcina mazei
brenda
Zhang, C.M.; Liu, C.; Slater, S.; Hou, Y.M.
Aminoacylation of tRNA with phosphoserine for synthesis of cysteinyl-tRNA(Cys)
Nat. Struct. Mol. Biol.
15
507-514
2008
Methanocaldococcus jannaschii
brenda
Fukunaga, R.; Harada, Y.; Hirao, I.; Yokoyama, S.
Phosphoserine aminoacylation of tRNA bearing an unnatural base anticodon
Biochem. Biophys. Res. Commun.
372
480-485
2008
Archaeoglobus fulgidus
brenda
Hauenstein, S.I.; Hou, Y.M.; Perona, J.J.
The homotetrameric phosphoseryl-tRNA synthetase from Methanosarcina mazei exhibits half-of-the-sites activity
J. Biol. Chem.
283
21997-22006
2008
Methanosarcina mazei
brenda
Kamtekar, S.; Hohn, M.J.; Park, H.S.; Schnitzbauer, M.; Sauerwald, A.; Sll, D.; Steitz, T.A.
Toward understanding phosphoseryl-tRNACys formation: the crystal structure of Methanococcus maripaludis phosphoseryl-tRNA synthetase
Proc. Natl. Acad. Sci. USA
104
2620-2625
2007
Methanococcus maripaludis (Q6LZE1), Methanococcus maripaludis
brenda
Englert, M.; Moses, S.; Hohn, M.; Ling, J.; ODonoghue, P.; Soell, D.
Aminoacylation of tRNA 2'- or 3'-hydroxyl by phosphoseryl- and pyrrolysyl-tRNA synthetases
FEBS Lett.
587
3360-3364
2013
Methanococcus maripaludis
brenda
Chen, M.; Nakazawa, Y.; Kubo, Y.; Asano, N.; Kato, K.; Tanaka, I.; Yao, M.
Crystallographic analysis of a subcomplex of the transsulfursome with tRNA for Cys-tRNACys synthesis
Acta Crystallogr. Sect. F
72
569-572
2016
Methanocaldococcus jannaschii (Q59054)
brenda
Mukai, T.; Crnkovic, A.; Umehara, T.; Ivanova, N.; Kyrpides, N.; Soll, D.
RNA-dependent cysteine biosynthesis in bacteria and archaea
mBio
8
e00561-17
2017
Archaeoglobus fulgidus, Methanococcus maripaludis, Candidatus Bathyarchaeota, Candidatus Parcubacteria, Chloroflexi
brenda
Chen, M.; Kato, K.; Kubo, Y.; Tanaka, Y.; Liu, Y.; Long, F.; Whitman, W.B.; Lill, P.; Gatsogiannis, C.; Raunser, S.; Shimizu, N.; Shinoda, A.; Nakamura, A.; Tanaka, I.; Yao, M.
Structural basis for tRNA-dependent cysteine biosynthesis
Nat. Commun.
8
1521
2017
Methanocaldococcus jannaschii (Q59054)
brenda
Liu, Y.; Nakamura, A.; Nakazawa, Y.; Asano, N.; Ford, K.A.; Hohn, M.J.; Tanaka, I.; Yao, M.; Soell, D.
Ancient translation factor is essential for tRNA-dependent cysteine biosynthesis in methanogenic archaea
Proc. Natl. Acad. Sci. USA
111
10520-10525
2014
Methanococcus maripaludis
brenda
Select items on the left to see more content.
EXTERNAL LINKS (specific for EC-Number 6.1.1.27)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
