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

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

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EC NUMBER	COMMENTARY
6.1.1.27	-

RECOMMENDED NAME	GeneOntology No.
O-phospho-L-serine-tRNA ligase	-

REACTION	REACTION DIAGRAM	COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
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		anticodon recognition mechanism	Archaeoglobus fulgidus	O30126	676139

REACTION TYPE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

PATHWAY	KEGG Link	MetaCyc Link
Aminoacyl-tRNA biosynthesis	00970	-
cysteine biosynthesis II (RNA-dependent)	-	PWY-6308

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].

SYNONYMS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
CysRS	Archaeoglobus fulgidus	O30126	-	676139
phosphoseryl-tRNA synthetase	Archaeoglobus fulgidus	-	-	690738
phosphoseryl-tRNA synthetase	Methanosarcina mazei	-	-	687743, 693106
phosphoseryl-tRNA synthetase	Methanococcus maripaludis	Q6LZE1	-	694852
phosphoseryl-tRNA synthetases	Archaeoglobus fulgidus	-	-	690738
SepRS	Methanocaldococcus jannaschii	-	SepRS–SepCysS binary complex	689145
SepRS	Archaeoglobus fulgidus	-	-	690738
SepRS	Methanosarcina mazei	-	-	687743, 693106
SepRS	Methanococcus maripaludis	-	-	684126, 694852
CysRS	Methanocaldococcus jannaschii	-	-	676139
additional information	Methanosarcina mazei	-	the enzyme is a class II tRNA synthetase and belongs to the PLP-dependent superfamily of enzymes	687743
additional information	Methanosarcina mazei	-	the enzyme is a class II tRNA synthetase	693106

CAS REGISTRY NUMBER	COMMENTARY
No entries in this field

ORGANISM	COMMENTARY	LITERATURE	SEQUENCE CODE	SEQUENCE DB	SOURCE
Archaeoglobus fulgidus	-	676139	O30126	SwissProt
Archaeoglobus fulgidus	-	690738	-	-
Methanocaldococcus jannaschii	-	676139, 689145	-	-
Methanococcus maripaludis	-	684126	-	-
Methanococcus maripaludis	-	694852	Q6LZE1	Uniprot
Methanosarcina mazei	-	687743, 693106	-	-
no activity in Methanobrevibacter smithii	-	684126	-	-
no activity in Methanosphaera stadtmanae	-	684126	-	-

GENERAL INFORMATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

SUBSTRATE	PRODUCT	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY/ Substrate	LITERATURE/ Substrate	COMMENTARY/ Product	LITERATURE/ Product	Reversibility r=reversible ir=irreversible ?=not specified
ATP + O-phospho-L-serine + tRNAAmber	AMP + diphosphate + O-phospho-L-serine-tRNAAmber		Archaeoglobus fulgidus	O30126	recognition of U34 and C35 of tRNAAmber by mutant E418N/E420N, no activity with wild-type SepRS, overview	676139	-	-	?
ATP + O-phospho-L-serine + tRNAAmber	AMP + diphosphate + O-phospho-L-seryl-tRNAAmber		Archaeoglobus fulgidus	-	mutant D418N/D420N/T423V	690738	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanocaldococcus jannaschii	-	-	689145	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Archaeoglobus fulgidus	-	-	690738	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanococcus maripaludis	-	-	684126	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	half-of-the-sites activity: the tetrameric enzyme binds two tRNAs. Only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate	693106	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanocaldococcus jannaschii	-	Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryl–tRNA synthetase (SepRS) acylates tRNACys with phosphoserine (Sep), and Sep-tRNA–Cys-tRNA synthase (SepCysS) converts the tRNA-bound phosphoserine to cysteine	689145	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanococcus maripaludis	-	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	684126	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Archaeoglobus fulgidus	-	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	690738	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	687743	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Archaeoglobus fulgidus	-	cognate substrate is tRNACys with the GCA anticodon, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon	690738	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	687743	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	693106	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	687743	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanocaldococcus jannaschii	-	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-Phosphoseryl–tRNA synthetase and Sep-tRNA–Cys-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, 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	689145	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	693106	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys		Methanocaldococcus jannaschii	-	-	676139	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys		Archaeoglobus fulgidus	O30126	-	676139	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys ?		Methanocaldococcus jannaschii	-	-	676139	-	-	?
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys ?		Archaeoglobus fulgidus	O30126	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	676139	-	-	?
ATP + O-phospho-L-serine + tRNAOpal	AMP + diphosphate + O-phospho-L-serine-tRNAOpal		Archaeoglobus fulgidus	O30126	recognition of U34 and C35 of tRNAOpal by mutant E418N/E420N, no activity with wild-type SepRS, overview	676139	-	-	?
ATP + O-phospho-L-serine + tRNAOpal	AMP + diphosphate + O-phospho-L-seryl-tRNAOpal		Archaeoglobus fulgidus	-	mutant D418N/D420N/T423V	690738	-	-	?
ATP + O-phospho-L-threonine + tRNACys	AMP + diphosphate + O-phospho-L-threonyl-tRNACys		Methanosarcina mazei	-	low activity, about 35% of the plateau aminoacylation observed with O-phospho-L-serine	693106	-	-	?
additional information	?	-	Archaeoglobus fulgidus	O30126	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	676139	-	-	-
additional information	?	-	Methanocaldococcus jannaschii	-	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	676139	-	-	-
additional information	?	-	Methanocaldococcus jannaschii	-	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, 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	689145	-	-	-
additional information	?	-	Methanosarcina mazei	-	SepRS is able to discriminate against the noncognate amino acids glutamate, serine, and phosphothreonine without the need for a separate hydrolytic editing site, 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	693106	-	-	-
additional information	?	-	Archaeoglobus fulgidus	-	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	690738	-	-	-
additional information	?	-	Archaeoglobus fulgidus	O30126	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	676139	-	-	-

NATURAL SUBSTRATES	NATURAL PRODUCTS	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY SUBSTRATE	LITERATURE (Substrate)	COMMENTARY PRODUCT	LITERATURE (Product)
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanocaldococcus jannaschii	-	-	689145	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Archaeoglobus fulgidus	-	-	690738	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	half-of-the-sites activity: the tetrameric enzyme binds two tRNAs. Only two of the four chemically equivalent subunits catalyze formation of phosphoseryl adenylate	693106	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanocaldococcus jannaschii	-	Methanocaldococcus jannaschii synthesizes Cys-tRNACys by an indirect pathway, whereby O-phosphoseryl–tRNA synthetase (SepRS) acylates tRNACys with phosphoserine (Sep), and Sep-tRNA–Cys-tRNA synthase (SepCysS) converts the tRNA-bound phosphoserine to cysteine	689145	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanococcus maripaludis	-	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	684126	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Archaeoglobus fulgidus	-	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	690738	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-seryl-tRNACys		Methanosarcina mazei	-	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	687743	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys		Methanocaldococcus jannaschii	-	-	676139	-	-
ATP + O-phospho-L-serine + tRNACys	AMP + diphosphate + O-phospho-L-serine-tRNACys		Archaeoglobus fulgidus	O30126	-	676139	-	-
additional information	?	-	Archaeoglobus fulgidus	O30126	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	676139	-	-
additional information	?	-	Methanocaldococcus jannaschii	-	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	676139	-	-
additional information	?	-	Methanocaldococcus jannaschii	-	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	689145	-	-

COFACTOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
ATP	Methanocaldococcus jannaschii	-	-	689145	2D-image
ATP	Archaeoglobus fulgidus	-	-	690738	2D-image
ATP	Methanosarcina mazei	-	-	687743, 693106	2D-image

METALS and IONS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
Mg2+	Methanocaldococcus jannaschii	-	-	689145
Mg2+	Archaeoglobus fulgidus	-	-	690738
Mg2+	Methanosarcina mazei	-	-	687743, 693106

INHIBITORS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

ACTIVATING COMPOUND	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

KM VALUE [mM]	KM VALUE [mM] Maximum	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
0.04	-	O-phospho-L-serine	Methanosarcina mazei	-	wild-type enzyme	693106	2D-image
0.15	-	O-phospho-L-serine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant mutant T307S	693106	2D-image
0.27	-	O-phospho-L-serine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	693106	2D-image
0.93	-	O-phospho-L-threonine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant mutant T307S	693106	2D-image
2.2	-	O-phospho-L-threonine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	693106	2D-image
8.4	-	O-phospho-L-threonine	Methanosarcina mazei	-	wild-type enzyme	693106	2D-image
0.0011	-	tRNACys	Methanocaldococcus jannaschii	-	60°C, pH 6.0, steady-state kinetics	689145	2D-image
0.0064	-	tRNACys	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	687743	2D-image
0.007	-	tRNACys	Methanocaldococcus jannaschii	-	pH 7.5, 37°C, recombinant enzyme, native tRNACys	689145	2D-image
0.0097	-	tRNACys	Methanocaldococcus jannaschii	-	pH 7.5, 37°C, recombinant enzyme, m1G37 tRNACys	689145	2D-image
0.0269	-	tRNACys	Archaeoglobus fulgidus	-	pH 7.6, 50°C, wild-type enzyme	690738	2D-image
0.0372	-	tRNACys	Archaeoglobus fulgidus	-	pH 7.6, 50°C, mutant D418N/D420N/T423V	690738	2D-image
0.151	-	tRNACys	Archaeoglobus fulgidus	-	50°C, pH 7.6, tRNACys containing the C(pyrrole-2-carbaldehyde)A anticodon, mutant enzyme D418N D420N T423V; 50°C, pH 7.6, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon, mutant enzyme D418N D420N T423V	690738	2D-image
0.269	-	tRNACys	Archaeoglobus fulgidus	-	50°C, pH 7.6, tRNACys with the GCA anticodon, wild-type enzyme	690738	2D-image
0.00097	-	m1G37-tRNACys	Methanocaldococcus jannaschii	-	60°C, pH 6.0, steady-state kinetics	689145	-
additional information	-	additional information	Methanosarcina mazei	-	Km-values for tRNACys isoacceptor; steady-state aminoacylation kinetics	687743	-
additional information	-	additional information	Methanocaldococcus jannaschii	-	steady-state and single-turnover kinetics, kinetic analysis, overview	689145	-
additional information	-	additional information	Archaeoglobus fulgidus	-	kinetic analysis of Sep-tRNA formation, overview	690738	-
additional information	-	additional information	Methanosarcina mazei	-	recombinant enzyme, ATP/diphosphate and aminoacylation kinetics, Michaelis-Menten kinetics	693106	-

TURNOVER NUMBER [1/s]	TURNOVER NUMBER MAXIMUM[1/s]	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
0.45	-	O-phospho-L-serine	Methanosarcina mazei	-	wild-type enzyme	693106	2D-image
10.6	-	O-phospho-L-serine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	693106	2D-image
11.2	-	O-phospho-L-serine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant mutant T307S	693106	2D-image
0.0054	-	O-phospho-L-threonine	Methanosarcina mazei	-	wild-type enzyme	693106	2D-image
0.014	-	O-phospho-L-threonine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	693106	2D-image
0.021	-	O-phospho-L-threonine	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant mutant T307S	693106	2D-image
0.008	-	tRNACys	Archaeoglobus fulgidus	-	pH 7.6, 50°C, mutant D418N/D420N/T423V	690738	2D-image
0.0471	-	tRNACys	Archaeoglobus fulgidus	-	50°C, pH 7.6, tRNACys containing the (pyrrole-2-carbaldehyde)UA anticodon, mutant enzyme D418N D420N T423V	690738	2D-image
0.048	-	tRNACys	Archaeoglobus fulgidus	-	50°C, pH 7.6, tRNACys containing the C(pyrrole-2-carbaldehyde)A anticodon, mutant enzyme D418N D420N T423V	690738	2D-image
0.07	-	tRNACys	Methanocaldococcus jannaschii	-	60°C, pH 6.0, steady-state kinetics	689145	2D-image
0.115	-	tRNACys	Archaeoglobus fulgidus	-	50°C, pH 7.6, tRNACys with the GCA anticodon, wild-type enzyme; pH 7.6, 50°C, wild-type enzyme	690738	2D-image
0.12	-	tRNACys	Methanosarcina mazei	-	pH 7.5, 37°C, recombinant enzyme	687743	2D-image
0.24	-	tRNACys	Methanocaldococcus jannaschii	-	pH 7.5, 37°C, recombinant enzyme, m1G37 tRNACys	689145	2D-image
1	-	tRNACys	Methanocaldococcus jannaschii	-	pH 7.5, 37°C, recombinant enzyme, native tRNACys	689145	2D-image
0.24	-	m1G37-tRNACys	Methanocaldococcus jannaschii	-	60°C, pH 6.0, steady-state kinetics	689145	-
additional information	-	additional information	Methanosarcina mazei	-	turnover number for tRNACys isoacceptor	687743	-

kcat/KM VALUE [1/mMs^-1]	kcat/KM VALUE [1/mMs^-1] Maximum	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

Ki VALUE [mM]	Ki VALUE [mM] Maximum	INHIBITOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

IC50 VALUE [mM]	IC50 VALUE [mM] Maximum	INHIBITOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

SPECIFIC ACTIVITY [µmol/min/mg]	SPECIFIC ACTIVITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
additional information	-	Methanosarcina mazei Goel, Methanosarcina mazei	-	recombinant enzyme, ATP-diphosphate exchange activity	693106

pH OPTIMUM	pH MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
7.5	-	Methanosarcina mazei	-	assay at	687743
7.5	-	Archaeoglobus fulgidus	-	assay at	690738
7.5	-	Methanosarcina mazei	-	assay at	693106
7.6	-	Archaeoglobus fulgidus	O30126	assay at	676139

pH RANGE	pH RANGE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

TEMPERATURE OPTIMUM	TEMPERATURE OPTIMUM MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
37	-	Methanosarcina mazei, Methanosarcina mazei Goel	-	assay at	687743
37	-	Methanosarcina mazei	-	assay at	693106
50	-	Archaeoglobus fulgidus	O30126	assay at	676139
50	-	Archaeoglobus fulgidus	-	assay at	690738

TEMPERATURE RANGE	TEMPERATURE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

pI VALUE	pI VALUE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

SOURCE TISSUE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	SOURCE
No entries in this field

LOCALIZATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	GeneOntology No.	LITERATURE	SOURCE
No entries in this field

PDB	SCOP	CATH	ORGANISM
No entries in this field

MOLECULAR WEIGHT	MOLECULAR WEIGHT MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
250000	-	Methanosarcina mazei	-	gel filtration	693106
250000	-	Methanosarcina mazei Goel	-	gel filtration; recombinant enzyme,native PAGE, and gel filtration	693106
250000	-	Methanosarcina mazei	-	recombinant enzyme,native PAGE, and gel filtration	693106
255400	-	Methanosarcina mazei	-	recombinant enzyme, mass spectrometry	693106

SUBUNITS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
tetramer	Archaeoglobus fulgidus	O30126	-	676139
tetramer	Methanocaldococcus jannaschii	-	-	676139
tetramer	Methanocaldococcus jannaschii	-	SepRS contains two tRNACys molecules per tetramer indicating an asymmetry of the four identical subunits	689145
tetramer	Methanosarcina mazei	-	4 * 60909, calculated from sequence; 4 * 60909, sequence calculation, 4 * 68992, homotetramer alpha4, mass spectrometry	693106
tetramer	Methanococcus maripaludis	Q6LZE1	homotetramer	694852
tetramer	Methanococcus maripaludis MM1265	-	homotetramer	694852
additional information	Methanocaldococcus jannaschii	-	both SepRS and SepCysS are active as a monomer in theSepRS–SepCysS binary complex	689145
additional information	Methanosarcina mazei	-	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	693106

POSTTRANSLATIONAL MODIFICATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

Crystallization/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	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	Archaeoglobus fulgidus	O30126	676139
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	Methanocaldococcus jannaschii	-	676139
hanging-drop vapor diffusion, 3.2 A resolution	Methanococcus maripaludis	Q6LZE1	694852
hanging-drop vapor diffusion, 3.2 A resolution	Methanococcus maripaludis MM1265	-	694852

pH STABILITY	pH STABILITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

TEMPERATURE STABILITY	TEMPERATURE STABILITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

GENERAL STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

ORGANIC SOLVENT	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

OXIDATION STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

STORAGE STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

Purification/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
recombinant	Archaeoglobus fulgidus	O30126	676139
recombinant	Methanocaldococcus jannaschii	-	676139
recombinant enzyme; recombinant His-tagged enzyme from Escherichia coli by anion exchange chromatography, gel filtration, and nickel affinity chromatography	Methanosarcina mazei	-	687743
recombinant; recombinant His-tagged wild-type and mutant enzymes from Escherichia coli to homogeneity by nickel affinity chromatography, the His-tag is cleaved off	Methanosarcina mazei	-	693106

Cloned/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), overexpression of SeMet-labeled SepRS in Escherichia coli strain B834(DE3)	Archaeoglobus fulgidus	O30126	676139
expression in Escherichia coli; expression of His-tagged enzyme in Escherichia coli	Methanosarcina mazei	-	687743
expression of His-tagged wild-type and mutant enzymes in Escherichia coli	Methanosarcina mazei	-	693106

EXPRESSION	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

ENGINEERING	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
D418N/D420N/T423V	Archaeoglobus fulgidus	-	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	690738
E418D	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418D/E420D	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418D/E420Q	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418N	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418N/E420D	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418N/E420N	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418N/E420N/T423V	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418N/E420N/T423V	Archaeoglobus fulgidus	-	efficiently charged phosphoserine to tRNA containing the (pyrrole-2-carbaldehyde)UA anticodon	690738
E418N/E420Q	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418Q	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418Q/E420D	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418Q/E420N	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E418Q/E420Q	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E420D	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E420K	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E420N	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
E420Q	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
T307S	Methanosarcina mazei	-	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	693106
E420R	Archaeoglobus fulgidus	O30126	site-directed mutagenesis, the mutant shows reduced activity and altered tRNA substrate specificity, compared to the wild-type enzyme	676139
additional information	Archaeoglobus fulgidus	O30126	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	676139
additional information	Methanocaldococcus jannaschii	-	mutation of the three anticodon nucleotides, G34, C35 and A36, as well as the next residue, G37, reduces the phosphoserylation activity	676139

Renatured/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

APPLICATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
biotechnology	Archaeoglobus fulgidus	O30126	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	676139

REF.	AUTHORS	TITLE	JOURNAL	VOL.	PAGES	YEAR	ORGANISM (UNIPROT ACCESSION NO.)	LINK TO PUBMED	SOURCE
676139	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	Archaeoglobus fulgidus (O30126), Methanocaldococcus jannaschii	PubMed
684126	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	PubMed
687743	Hauenstein, S.I.; Perona, J.J.	Redundant synthesis of cysteinyl-tRNACys in Methanosarcina mazei	J. Biol. Chem.	283	22007-22017	2008	Methanosarcina mazei, Methanosarcina mazei (Q8PVS9)	PubMed
689145	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, Methanocaldococcus jannaschii (Q59072)	PubMed
690738	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	PubMed
693106	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	PubMed
694852	Kamtekar, S.; Hohn, M.J.; Park, H.S.; Schnitzbauer, M.; Sauerwald, A.; Söll, 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, Methanococcus maripaludis (Q6LZE1)	PubMed

LINKS TO OTHER DATABASES (specific for EC-Number 6.1.1.27)

NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM

IUBMB Enzyme Nomenclature

PROSITE Database of protein families and domains

SYSTERS

Protein Mutant Database

InterPro (database of protein families, domains and functional sites)

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