SYN_ECOLI
ID SYN_ECOLI Reviewed; 466 AA.
AC P0A8M0; P17242;
DT 07-JUN-2005, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 03-AUG-2022, entry version 131.
DE RecName: Full=Asparagine--tRNA ligase;
DE EC=6.1.1.22 {ECO:0000269|PubMed:1544480, ECO:0000269|PubMed:2009959};
DE AltName: Full=Asparaginyl-tRNA synthetase;
DE Short=AsnRS;
GN Name=asnS; Synonyms=tss; OrderedLocusNames=b0930, JW0913;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-12 AND
RP 243-256.
RC STRAIN=CE1215;
RX PubMed=2693216; DOI=10.1016/0378-1119(89)90524-6;
RA Anselme J., Haertlein M.;
RT "Asparaginyl-tRNA synthetase from Escherichia coli has significant sequence
RT homologies with yeast aspartyl-tRNA synthetase.";
RL Gene 84:481-485(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=1425658; DOI=10.1111/j.1432-1033.1992.tb17315.x;
RA Aoki H., Yaworsky P.J., Patel S.D., Margolin-Brzezinski D., Park K.S.,
RA Ganoza M.C.;
RT "The asparaginyl-tRNA synthetase gene encodes one of the complementing
RT factors for thermosensitive translation in the Escherichia coli mutant
RT strain, N4316.";
RL Eur. J. Biochem. 209:511-521(1992).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=8905232; DOI=10.1093/dnares/3.3.137;
RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K.,
RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S.,
RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H.,
RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G.,
RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M.,
RA Horiuchi T.;
RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 12.7-28.0 min region on the linkage map.";
RL DNA Res. 3:137-155(1996).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [6]
RP PROTEIN SEQUENCE OF 2-13.
RC STRAIN=K12 / EMG2;
RX PubMed=9298646; DOI=10.1002/elps.1150180807;
RA Link A.J., Robison K., Church G.M.;
RT "Comparing the predicted and observed properties of proteins encoded in the
RT genome of Escherichia coli K-12.";
RL Electrophoresis 18:1259-1313(1997).
RN [7]
RP ATP-BINDING SITE, MUTAGENESIS OF TYR-426, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=2009959; DOI=10.1016/0014-5793(91)80228-u;
RA Anselme J., Haertlein M.;
RT "Tyr-426 of the Escherichia coli asparaginyl-tRNA synthetase, an amino acid
RT in a C-terminal conserved motif, is involved in ATP binding.";
RL FEBS Lett. 280:163-166(1991).
RN [8]
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTANT
RP TEMPERATURE-SENSITIVE HO202.
RC STRAIN=K12, and K12 / TsHO202;
RX PubMed=1544480; DOI=10.1016/0014-5793(92)80106-q;
RA Madern D., Anselme J., Haertlein M.;
RT "Asparaginyl-tRNA synthetase from the Escherichia coli temperature-
RT sensitive strain HO202. A proline replacement in motif 2 is responsible for
RT a large increase in Km for asparagine and ATP.";
RL FEBS Lett. 299:85-89(1992).
RN [9]
RP IDENTIFICATION BY 2D-GEL.
RX PubMed=9298644; DOI=10.1002/elps.1150180805;
RA VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.;
RT "Escherichia coli proteome analysis using the gene-protein database.";
RL Electrophoresis 18:1243-1251(1997).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-asparagine + tRNA(Asn) = AMP + diphosphate + H(+) + L-
CC asparaginyl-tRNA(Asn); Xref=Rhea:RHEA:11180, Rhea:RHEA-COMP:9659,
CC Rhea:RHEA-COMP:9674, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:58048, ChEBI:CHEBI:78442,
CC ChEBI:CHEBI:78515, ChEBI:CHEBI:456215; EC=6.1.1.22;
CC Evidence={ECO:0000269|PubMed:1544480, ECO:0000269|PubMed:2009959};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=29 uM for asparagine {ECO:0000269|PubMed:1544480};
CC KM=15.3 uM for asparagine {ECO:0000269|PubMed:2009959};
CC KM=0.5 mM for ATP {ECO:0000269|PubMed:2009959};
CC -!- SUBUNIT: Homodimer.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- SIMILARITY: Belongs to the class-II aminoacyl-tRNA synthetase family.
CC {ECO:0000305}.
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DR EMBL; M33145; AAA24666.1; -; Genomic_DNA.
DR EMBL; X68192; CAA48274.1; -; Genomic_DNA.
DR EMBL; U00096; AAC74016.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35682.1; -; Genomic_DNA.
DR PIR; JS0396; SYECNT.
DR RefSeq; NP_415450.1; NC_000913.3.
DR RefSeq; WP_000117881.1; NZ_STEB01000006.1.
DR AlphaFoldDB; P0A8M0; -.
DR SMR; P0A8M0; -.
DR BioGRID; 4259395; 8.
DR DIP; DIP-9179N; -.
DR IntAct; P0A8M0; 3.
DR STRING; 511145.b0930; -.
DR SWISS-2DPAGE; P0A8M0; -.
DR jPOST; P0A8M0; -.
DR PaxDb; P0A8M0; -.
DR PRIDE; P0A8M0; -.
DR EnsemblBacteria; AAC74016; AAC74016; b0930.
DR EnsemblBacteria; BAA35682; BAA35682; BAA35682.
DR GeneID; 66670794; -.
DR GeneID; 945555; -.
DR KEGG; ecj:JW0913; -.
DR KEGG; eco:b0930; -.
DR PATRIC; fig|1411691.4.peg.1345; -.
DR EchoBASE; EB0092; -.
DR eggNOG; COG0017; Bacteria.
DR HOGENOM; CLU_004553_2_0_6; -.
DR InParanoid; P0A8M0; -.
DR OMA; DNMDLAE; -.
DR PhylomeDB; P0A8M0; -.
DR BioCyc; EcoCyc:ASNS-MON; -.
DR BioCyc; MetaCyc:ASNS-MON; -.
DR PRO; PR:P0A8M0; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004816; F:asparagine-tRNA ligase activity; IDA:EcoCyc.
DR GO; GO:0005524; F:ATP binding; IMP:EcoCyc.
DR GO; GO:0003676; F:nucleic acid binding; IEA:InterPro.
DR GO; GO:0006421; P:asparaginyl-tRNA aminoacylation; IDA:EcoCyc.
DR GO; GO:0006418; P:tRNA aminoacylation for protein translation; IDA:EcoCyc.
DR Gene3D; 2.40.50.140; -; 1.
DR Gene3D; 3.30.930.10; -; 1.
DR HAMAP; MF_00534; Asn_tRNA_synth; 1.
DR InterPro; IPR004364; Aa-tRNA-synt_II.
DR InterPro; IPR006195; aa-tRNA-synth_II.
DR InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL.
DR InterPro; IPR004522; Asn-tRNA-ligase.
DR InterPro; IPR002312; Asp/Asn-tRNA-synth_IIb.
DR InterPro; IPR012340; NA-bd_OB-fold.
DR InterPro; IPR004365; NA-bd_OB_tRNA.
DR Pfam; PF00152; tRNA-synt_2; 1.
DR Pfam; PF01336; tRNA_anti-codon; 1.
DR PRINTS; PR01042; TRNASYNTHASP.
DR SUPFAM; SSF50249; SSF50249; 1.
DR SUPFAM; SSF55681; SSF55681; 1.
DR TIGRFAMs; TIGR00457; asnS; 1.
DR PROSITE; PS50862; AA_TRNA_LIGASE_II; 1.
PE 1: Evidence at protein level;
KW Aminoacyl-tRNA synthetase; ATP-binding; Cytoplasm;
KW Direct protein sequencing; Ligase; Nucleotide-binding;
KW Protein biosynthesis; Reference proteome.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:2693216,
FT ECO:0000269|PubMed:9298646"
FT CHAIN 2..466
FT /note="Asparagine--tRNA ligase"
FT /id="PRO_0000176406"
FT VARIANT 231
FT /note="P -> L (in temperature-sensitive mutant HO202)"
FT MUTAGEN 426
FT /note="Y->F: No effect."
FT /evidence="ECO:0000269|PubMed:2009959"
FT MUTAGEN 426
FT /note="Y->S: 15-fold increase in Km for ATP."
FT /evidence="ECO:0000269|PubMed:2009959"
SQ SEQUENCE 466 AA; 52570 MW; 1E477CB5467B772F CRC64;
MSVVPVADVL QGRVAVDSEV TVRGWVRTRR DSKAGISFLA VYDGSCFDPV QAVINNSLPN
YNEDVLRLTT GCSVIVTGKV VASPGQGQQF EIQASKVEVA GWVEDPDTYP MAAKRHSIEY
LREVAHLRPR TNLIGAVARV RHTLAQALHR FFNEQGFFWV STPLITASDT EGAGEMFRVS
TLDLENLPRN DQGKVDFDKD FFGKESFLTV SGQLNGETYA CALSKIYTFG PTFRAENSNT
SRHLAEFWML EPEVAFANLN DIAGLAEAML KYVFKAVLEE RADDMKFFAE RVDKDAVSRL
ERFIEADFAQ VDYTDAVTIL ENCGRKFENP VYWGVDLSSE HERYLAEEHF KAPVVVKNYP
KDIKAFYMRL NEDGKTVAAM DVLAPGIGEI IGGSQREERL DVLDERMLEM GLNKEDYWWY
RDLRRYGTVP HSGFGLGFER LIAYVTGVQN VRDVIPFPRT PRNASF
