ID   SYA_ECOLI               Reviewed;         876 AA.
AC   P00957; P78279;
DT   21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1997, sequence version 2.
DT   04-NOV-2026, entry version 222.
DE   RecName: Full=Alanine--tRNA ligase;
DE            EC=6.1.1.7 {ECO:0000269|PubMed:30274179, ECO:0000269|PubMed:7005211};
DE   AltName: Full=Alanyl-tRNA synthetase;
DE            Short=AlaRS;
DE   AltName: Full=Protein lactyltransferase {ECO:0000305};
DE            EC=6.-.-.- {ECO:0000269|PubMed:38653238, ECO:0000269|PubMed:39322678};
GN   Name=alaS; Synonyms=lovB; OrderedLocusNames=b2697, JW2667;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Enterobacterales; Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=9205837; DOI=10.1093/dnares/4.2.91;
RA   Yamamoto Y., Aiba H., Baba T., Hayashi K., Inada T., Isono K., Itoh T.,
RA   Kimura S., Kitagawa M., Makino K., Miki T., Mitsuhashi N., Mizobuchi K.,
RA   Mori H., Nakade S., Nakamura Y., Nashimoto H., Oshima T., Oyama S.,
RA   Saito N., Sampei G., Satoh Y., Sivasundaram S., Tagami H., Takahashi H.,
RA   Takeda J., Takemoto K., Uehara K., Wada C., Yamagata S., Horiuchi T.;
RT   "Construction of a contiguous 874-kb sequence of the Escherichia coli-K12
RT   genome corresponding to 50.0-68.8 min on the linkage map and analysis of
RT   its sequence features.";
RL   DNA Res. 4:91-113(1997).
RN   [2]
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   [3]
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   [4]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-166, AND PARTIAL PROTEIN SEQUENCE.
RX   PubMed=7005898; DOI=10.1073/pnas.77.11.6531;
RA   Herlihy W.C., Royal N.J., Biemann K., Putney S.D., Schimmel P.R.;
RT   "Mass spectra of partial protein hydrolysates as a multiple phase check for
RT   long polypeptides deduced from DNA sequences: NH2-terminal segment of
RT   alanine tRNA synthetase.";
RL   Proc. Natl. Acad. Sci. U.S.A. 77:6531-6535(1980).
RN   [5]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 2-876.
RX   PubMed=7025207; DOI=10.1126/science.7025207;
RA   Putney S.D., Royal N.J., de Vegvar H.N., Herlihy W.C., Biemann K.,
RA   Schimmel P.;
RT   "Primary structure of a large aminoacyl-tRNA synthetase.";
RL   Science 213:1497-1501(1981).
RN   [6]
RP   PROTEIN SEQUENCE OF 2-12, SUBUNIT, AND CATALYTIC ACTIVITY.
RX   PubMed=7005211; DOI=10.1016/s0021-9258(19)70119-7;
RA   Putney S.D., Sauer R.T., Schimmel P.R.;
RT   "Purification and properties of alanine tRNA synthetase from Escherichia
RT   coli A tetramer of identical subunits.";
RL   J. Biol. Chem. 256:198-204(1981).
RN   [7]
RP   PROTEIN SEQUENCE OF 163-173.
RX   PubMed=8292605; DOI=10.1021/bi00169a019;
RA   Musier-Forsyth K., Schimmel P.;
RT   "Acceptor helix interactions in a class II tRNA synthetase: photoaffinity
RT   cross-linking of an RNA miniduplex substrate.";
RL   Biochemistry 33:773-779(1994).
RN   [8]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 799-876.
RC   STRAIN=KP4714;
RX   PubMed=8604133; DOI=10.1006/jmbi.1996.0103;
RA   Murayama N., Shimizu H., Takiguchi S., Baba Y., Amino H., Horiuchi T.,
RA   Sekimizu K., Miki T.;
RT   "Evidence for involvement of Escherichia coli genes pmbA, csrA and a
RT   previously unrecognized gene tldD, in the control of DNA gyrase by letD
RT   (ccdB) of sex factor F.";
RL   J. Mol. Biol. 256:483-502(1996).
RN   [9]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 866-876.
RC   STRAIN=K12;
RX   PubMed=8393005; DOI=10.1128/jb.175.15.4744-4755.1993;
RA   Romeo T., Gong M., Liu M.-Y., Brun-Zinkernagel A.-M.;
RT   "Identification and molecular characterization of csrA, a pleiotropic gene
RT   from Escherichia coli that affects glycogen biosynthesis, gluconeogenesis,
RT   cell size, and surface properties.";
RL   J. Bacteriol. 175:4744-4755(1993).
RN   [10]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-13.
RX   PubMed=6256345; DOI=10.1016/s0021-9258(19)70120-3;
RA   Putney S.D., Melendez D.L., Schimmel P.R.;
RT   "Cloning, partial sequencing, and in vitro transcription of the gene for
RT   alanine tRNA synthetase.";
RL   J. Biol. Chem. 256:205-211(1981).
RN   [11]
RP   ZINC-BINDING, AND MUTAGENESIS OF CYS-179; CYS-182; GLU-184; ASP-188;
RP   HIS-189; ASP-191 AND HIS-192.
RX   PubMed=1712632; DOI=10.1021/bi00242a023;
RA   Miller W.T., Hill K.A.W., Schimmel P.;
RT   "Evidence for a 'cysteine-histidine box' metal-binding site in an
RT   Escherichia coli aminoacyl-tRNA synthetase.";
RL   Biochemistry 30:6970-6976(1991).
RN   [12]
RP   FUNCTION IN AMINOACYLATION OF TMRNA.
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=7524073; DOI=10.1073/pnas.91.20.9223;
RA   Komine Y., Kitabatake M., Yokogawa T., Nishikawa K., Inokuchi H.;
RT   "A tRNA-like structure is present in 10Sa RNA, a small stable RNA from
RT   Escherichia coli.";
RL   Proc. Natl. Acad. Sci. U.S.A. 91:9223-9227(1994).
RN   [13]
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).
RN   [14]
RP   FUNCTION IN TRNA EDITING, AND MUTAGENESIS OF HIS-564; HIS-568; GLN-584;
RP   CYS-666 AND HIS-670.
RX   PubMed=12554667; DOI=10.1093/emboj/cdg065;
RA   Beebe K., Ribas De Pouplana L., Schimmel P.;
RT   "Elucidation of tRNA-dependent editing by a class II tRNA synthetase and
RT   significance for cell viability.";
RL   EMBO J. 22:668-675(2003).
RN   [15]
RP   FUNCTION IN TRNA EDITING, AND MUTAGENESIS OF GLN-584 AND CYS-666.
RC   STRAIN=K12;
RX   PubMed=18723508; DOI=10.1074/jbc.m805943200;
RA   Chong Y.E., Yang X.L., Schimmel P.;
RT   "Natural homolog of tRNA synthetase editing domain rescues conditional
RT   lethality caused by mistranslation.";
RL   J. Biol. Chem. 283:30073-30078(2008).
RN   [16]
RP   DOMAIN EDITING, AND MUTAGENESIS OF ARG-693.
RX   PubMed=18172502; DOI=10.1038/nature06454;
RA   Beebe K., Mock M., Merriman E., Schimmel P.;
RT   "Distinct domains of tRNA synthetase recognize the same base pair.";
RL   Nature 451:90-93(2008).
RN   [17]
RP   ACETYLATION [LARGE SCALE ANALYSIS] AT LYS-74, AND IDENTIFICATION BY MASS
RP   SPECTROMETRY.
RC   STRAIN=K12 / JW1106, and K12 / MG1655 / ATCC 47076;
RX   PubMed=18723842; DOI=10.1074/mcp.m800187-mcp200;
RA   Zhang J., Sprung R., Pei J., Tan X., Kim S., Zhu H., Liu C.F.,
RA   Grishin N.V., Zhao Y.;
RT   "Lysine acetylation is a highly abundant and evolutionarily conserved
RT   modification in Escherichia coli.";
RL   Mol. Cell. Proteomics 8:215-225(2009).
RN   [18]
RP   FUNCTION IN TRNA-BINDING VIA C-ALA DOMAIN, AND DOMAIN EXCHANGE EXPERIMENTS.
RX   PubMed=19661429; DOI=10.1126/science.1174343;
RA   Guo M., Chong Y.E., Beebe K., Shapiro R., Yang X.-L., Schimmel P.;
RT   "The C-Ala domain brings together editing and aminoacylation functions on
RT   one tRNA.";
RL   Science 325:744-747(2009).
RN   [19]
RP   FUNCTION, AND MUTAGENESIS OF THR-567; SER-587 AND CYS-666.
RX   PubMed=28362257; DOI=10.7554/elife.24001;
RA   Pawar K.I., Suma K., Seenivasan A., Kuncha S.K., Routh S.B.,
RA   Kruparani S.P., Sankaranarayanan R.;
RT   "Role of D-aminoacyl-tRNA deacylase beyond chiral proofreading as a
RT   cellular defense against glycine mischarging by AlaRS.";
RL   Elife 6:0-0(2017).
RN   [20]
RP   FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, ACETYLATION AT LYS-74,
RP   AND MUTAGENESIS OF LYS-74.
RC   STRAIN=K12 / DH10B;
RX   PubMed=30274179; DOI=10.3390/genes9100473;
RA   Umehara T., Kosono S., Soell D., Tamura K.;
RT   "Lysine Acetylation Regulates Alanyl-tRNA Synthetase Activity in
RT   Escherichia coli.";
RL   Genes (Basel) 9:0-0(2018).
RN   [21]
RP   FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF MET-44; ARG-70; TRP-171;
RP   ASN-213; 236-ASP--GLY-238 AND GLY-240.
RX   PubMed=39322678; DOI=10.1038/s41586-024-07992-y;
RA   Li H., Liu C., Li R., Zhou L., Ran Y., Yang Q., Huang H., Lu H., Song H.,
RA   Yang B., Ru H., Lin S., Zhang L.;
RT   "AARS1 and AARS2 sense L-lactate to regulate cGAS as global lysine
RT   lactyltransferases.";
RL   Nature 0:0-0(2024).
RN   [22]
RP   FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF MET-44; ARG-70; TRP-171;
RP   ASN-213 AND 236-ASP--GLY-238.
RX   PubMed=38653238; DOI=10.1016/j.cell.2024.04.002;
RA   Zong Z., Xie F., Wang S., Wu X., Zhang Z., Yang B., Zhou F.;
RT   "Alanyl-tRNA synthetase, AARS1, is a lactate sensor and lactyltransferase
RT   that lactylates p53 and contributes to tumorigenesis.";
RL   Cell 187:2375-2392(2024).
RN   [23] {ECO:0007744|PDB:3HXU, ECO:0007744|PDB:3HXV, ECO:0007744|PDB:3HXW, ECO:0007744|PDB:3HXX, ECO:0007744|PDB:3HXY, ECO:0007744|PDB:3HXZ, ECO:0007744|PDB:3HY0, ECO:0007744|PDB:3HY1}
RP   X-RAY CRYSTALLOGRAPHY WITH AND WITHOUT CHARGED CORRECT AND INCORRECT AMINO
RP   ACID, AND MUTAGENESIS OF ASP-236 AND GLY-238.
RX   PubMed=20010690; DOI=10.1038/nature08612;
RA   Guo M., Chong Y.E., Shapiro R., Beebe K., Yang X.L., Schimmel P.;
RT   "Paradox of mistranslation of serine for alanine caused by AlaRS
RT   recognition dilemma.";
RL   Nature 462:808-812(2009).
CC   -!- FUNCTION: Catalyzes the attachment of L-alanine to tRNA(Ala) in a two-
CC       step reaction: L-alanine is first activated by ATP to form Ala-AMP and
CC       then transferred to the acceptor end of tRNA(Ala) (PubMed:28362257).
CC       AlaRS also incorrectly activates the sterically smaller amino acid
CC       glycine as well as the sterically larger amino acid L-serine; generates
CC       2-fold more mischarged Gly than Ser (PubMed:28362257). These mischarged
CC       amino acids occur because the of inherent physicochemical limitations
CC       on discrimination between closely related amino acids (Ala, Gly and
CC       Ser) in the charging step (PubMed:28362257). In presence of high levels
CC       of lactate, also acts as a protein lactyltransferase that mediates
CC       lactylation of lysine residues in target proteins (PubMed:38653238,
CC       PubMed:39322678). {ECO:0000269|PubMed:28362257,
CC       ECO:0000269|PubMed:38653238, ECO:0000269|PubMed:39322678}.
CC   -!- FUNCTION: Edits mischarged Ser-tRNA(Ala) and Gly-tRNA(Ala) but not
CC       incorrectly charged Ser-tRNA(Thr) (PubMed:12554667, PubMed:18723508).
CC       Dtd edits Gly-tRNA(Ala) 4-fold better than does AlaRS
CC       (PubMed:28362257). {ECO:0000269|PubMed:12554667,
CC       ECO:0000269|PubMed:18723508, ECO:0000269|PubMed:28362257}.
CC   -!- FUNCTION: Attaches Ala to transfer-messenger RNA (tmRNA, also known as
CC       10Sa RNA, the product of the ssrA gene). tmRNA plays a major role in
CC       rescue of stalled ribosomes via trans-translation.
CC       {ECO:0000269|PubMed:7524073}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=tRNA(Ala) + L-alanine + ATP = L-alanyl-tRNA(Ala) + AMP +
CC         diphosphate; Xref=Rhea:RHEA:12540, Rhea:RHEA-COMP:9657, Rhea:RHEA-
CC         COMP:9923, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:57972,
CC         ChEBI:CHEBI:78442, ChEBI:CHEBI:78497, ChEBI:CHEBI:456215; EC=6.1.1.7;
CC         Evidence={ECO:0000269|PubMed:30274179, ECO:0000269|PubMed:7005211};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:12541;
CC         Evidence={ECO:0000305|PubMed:7005211};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-lactate + ATP + H(+) = (S)-lactoyl-AMP + diphosphate;
CC         Xref=Rhea:RHEA:80271, ChEBI:CHEBI:15378, ChEBI:CHEBI:16651,
CC         ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:231470;
CC         Evidence={ECO:0000269|PubMed:38653238};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:80272;
CC         Evidence={ECO:0000269|PubMed:38653238};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-lactoyl-AMP + L-lysyl-[protein] = N(6)-[(S)-lactoyl]-L-
CC         lysyl-[protein] + AMP + 2 H(+); Xref=Rhea:RHEA:80275, Rhea:RHEA-
CC         COMP:9752, Rhea:RHEA-COMP:19466, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:29969, ChEBI:CHEBI:231470, ChEBI:CHEBI:231527,
CC         ChEBI:CHEBI:456215; Evidence={ECO:0000269|PubMed:38653238};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:80276;
CC         Evidence={ECO:0000269|PubMed:38653238};
CC   -!- COFACTOR:
CC       Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC       Note=Binds 1 zinc ion per subunit; it is not clear where this binding
CC       occurs. {ECO:0000269|PubMed:1712632};
CC   -!- ACTIVITY REGULATION: Acetylation at Lys-74 decreases the alanylation
CC       activity for tRNA(Ala); a protein that is fully acetylated is inactive
CC       in vitro. {ECO:0000269|PubMed:30274179}.
CC   -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:7005211}.
CC   -!- INTERACTION:
CC       P00957; P00957: alaS; NbExp=2; IntAct=EBI-544061, EBI-544061;
CC   -!- SUBCELLULAR LOCATION: Cytoplasm.
CC   -!- DOMAIN: Consists of three domains; the N-terminal catalytic domain, the
CC       editing domain and the C-terminal C-Ala domain. The editing domain
CC       removes incorrectly charged amino acids, while the C-Ala domain, along
CC       with tRNA(Ala), serves as a bridge to cooperatively bring together the
CC       editing and aminoacylation centers thus stimulating deacylation of
CC       misacylated tRNAs.
CC   -!- DOMAIN: The editing domain removes incorrectly charged amino acids,
CC       i.e. Ser-tRNA(Ala) or Gly-tRNA(Ala) become uncharged tRNA(Ala) and the
CC       amino acid. It is specific for the acceptor stem of tRNA(Ala).
CC   -!- DOMAIN: The C-terminal C-Ala domain, along with tRNA(Ala), serves as a
CC       bridge to cooperatively bring together the editing and aminoacylation
CC       centers thus stimulating deacylation of misacylated tRNAs. This C-Ala
CC       domain can be replaced in vitro by the corresponding domain of Aquifex
CC       aeolicus or man. Recognition of tRNA(Ala) by the 2 domains is
CC       independent, that is one enzyme recognizes the same tRNA(Ala) in 2
CC       different manners. {ECO:0000305}.
CC   -!- SIMILARITY: Belongs to the class-II aminoacyl-tRNA synthetase family.
CC       {ECO:0000305}.
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DR   EMBL; U00096; AAC75739.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAA16559.1; -; Genomic_DNA.
DR   EMBL; J01581; AAA03208.1; -; Unassigned_DNA.
DR   EMBL; L07596; AAA71918.1; ALT_SEQ; Unassigned_DNA.
DR   EMBL; D44453; BAA21554.1; -; Genomic_DNA.
DR   EMBL; Z28405; CAA82247.1; -; Genomic_DNA.
DR   PIR; E65049; SYECAT.
DR   RefSeq; NP_417177.1; NC_000913.3.
DR   RefSeq; WP_000047184.1; NZ_CP009789.1.
DR   PDB; 3HXU; X-ray; 2.10 A; A=2-442.
DR   PDB; 3HXV; X-ray; 1.93 A; A=2-442.
DR   PDB; 3HXW; X-ray; 1.93 A; A=2-442.
DR   PDB; 3HXX; X-ray; 2.11 A; A=2-442.
DR   PDB; 3HXY; X-ray; 2.27 A; A=2-442.
DR   PDB; 3HXZ; X-ray; 1.99 A; A/B/C/D=2-442.
DR   PDB; 3HY0; X-ray; 1.90 A; A/B=2-442.
DR   PDB; 3HY1; X-ray; 2.79 A; A/B=2-442.
DR   AlphaFoldDB; P00957; -.
DR   MDposit; P00957; -.
DR   SMR; P00957; -.
DR   BioGRID; 4261283; 66.
DR   BioGRID; 851507; 1.
DR   DIP; DIP-9080N; -.
DR   FunCoup; P00957; 916.
DR   IntAct; P00957; 22.
DR   STRING; 511145.b2697; -.
DR   MoonProt; P00957; -.
DR   iPTMnet; P00957; -.
DR   jPOST; P00957; -.
DR   PaxDb; 511145-b2697; -.
DR   EnsemblBacteria; AAC75739; AAC75739; b2697.
DR   GeneID; 947175; -.
DR   KEGG; ecj:JW2667; -.
DR   KEGG; eco:b2697; -.
DR   KEGG; ecoc:C3026_14845; -.
DR   PATRIC; fig|1411691.4.peg.4046; -.
DR   EchoBASE; EB0033; -.
DR   eggNOG; COG0013; Bacteria.
DR   HOGENOM; CLU_004485_1_1_6; -.
DR   InParanoid; P00957; -.
DR   OMA; AAAKWMY; -.
DR   OrthoDB; 9803884at2; -.
DR   PhylomeDB; P00957; -.
DR   BioCyc; EcoCyc:ALAS-MONOMER; -.
DR   BioCyc; MetaCyc:ALAS-MONOMER; -.
DR   BRENDA; 6.1.1.7; 2026.
DR   EvolutionaryTrace; P00957; -.
DR   PRO; PR:P00957; -.
DR   Proteomes; UP000000625; Chromosome.
DR   AbasyAtlas; P00957; -.
DR   GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR   GO; GO:0016020; C:membrane; HDA:UniProtKB.
DR   GO; GO:0004813; F:alanine-tRNA ligase activity; IDA:EcoCyc.
DR   GO; GO:0002161; F:aminoacyl-tRNA deacylase activity; IDA:UniProtKB.
DR   GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR   GO; GO:0001217; F:DNA-binding transcription repressor activity; IDA:EcoCyc.
DR   GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR   GO; GO:0003676; F:nucleic acid binding; IEA:InterPro.
DR   GO; GO:0141207; F:peptide lactyltransferase (ATP-dependent) activity; IDA:UniProtKB.
DR   GO; GO:0042803; F:protein homodimerization activity; IDA:EcoCyc.
DR   GO; GO:0002196; F:Ser-tRNA(Ala) deacylase activity; IDA:EcoCyc.
DR   GO; GO:0008270; F:zinc ion binding; IDA:EcoCyc.
DR   GO; GO:0006419; P:alanyl-tRNA aminoacylation; IDA:EcoCyc.
DR   GO; GO:0045892; P:negative regulation of DNA-templated transcription; IDA:EcoCyc.
DR   CDD; cd00673; AlaRS_core; 1.
DR   FunFam; 2.40.30.130:FF:000001; Alanine--tRNA ligase; 1.
DR   FunFam; 3.10.310.40:FF:000001; Alanine--tRNA ligase; 1.
DR   FunFam; 3.30.930.10:FF:000004; Alanine--tRNA ligase; 1.
DR   FunFam; 3.30.980.10:FF:000004; Alanine--tRNA ligase, cytoplasmic; 1.
DR   Gene3D; 2.40.30.130; -; 1.
DR   Gene3D; 3.10.310.40; -; 1.
DR   Gene3D; 3.30.54.20; -; 1.
DR   Gene3D; 6.10.250.550; -; 1.
DR   Gene3D; 3.30.930.10; Bira Bifunctional Protein, Domain 2; 1.
DR   Gene3D; 3.30.980.10; Threonyl-trna Synthetase, Chain A, domain 2; 1.
DR   HAMAP; MF_00036_B; Ala_tRNA_synth_B; 1.
DR   InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL.
DR   InterPro; IPR002318; Ala-tRNA-lgiase_IIc.
DR   InterPro; IPR018162; Ala-tRNA-ligase_IIc_anticod-bd.
DR   InterPro; IPR018165; Ala-tRNA-synth_IIc_core.
DR   InterPro; IPR018164; Ala-tRNA-synth_IIc_N.
DR   InterPro; IPR023033; Ala_tRNA_ligase_euk/bac.
DR   InterPro; IPR003156; DHHA1_dom.
DR   InterPro; IPR018163; Thr/Ala-tRNA-synth_IIc_edit.
DR   InterPro; IPR009000; Transl_B-barrel_sf.
DR   InterPro; IPR012947; tRNA_SAD.
DR   NCBIfam; TIGR00344; alaS; 1.
DR   PANTHER; PTHR11777:SF9; ALANINE--TRNA LIGASE, CYTOPLASMIC; 1.
DR   PANTHER; PTHR11777; ALANYL-TRNA SYNTHETASE; 1.
DR   Pfam; PF02272; DHHA1; 1.
DR   Pfam; PF01411; tRNA-synt_2c; 1.
DR   Pfam; PF07973; tRNA_SAD; 1.
DR   PRINTS; PR00980; TRNASYNTHALA.
DR   SMART; SM00863; tRNA_SAD; 1.
DR   SUPFAM; SSF55681; Class II aaRS and biotin synthetases; 1.
DR   SUPFAM; SSF101353; Putative anticodon-binding domain of alanyl-tRNA synthetase (AlaRS); 1.
DR   SUPFAM; SSF55186; ThrRS/AlaRS common domain; 1.
DR   SUPFAM; SSF50447; Translation proteins; 1.
DR   PROSITE; PS50860; AA_TRNA_LIGASE_II_ALA; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Acetylation; Aminoacyl-tRNA synthetase; ATP-binding;
KW   Cytoplasm; Direct protein sequencing; Ligase; Metal-binding;
KW   Nucleotide-binding; Protein biosynthesis; Reference proteome; RNA-binding;
KW   tRNA-binding; Zinc.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:7005211"
FT   CHAIN           2..876
FT                   /note="Alanine--tRNA ligase"
FT                   /id="PRO_0000075113"
FT   REGION          2..461
FT                   /note="Catalytic"
FT   REGION          553..705
FT                   /note="Editing"
FT   REGION          699..808
FT                   /note="Important for oligomerization"
FT   REGION          766..875
FT                   /note="C-Ala domain"
FT   BINDING         564
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /evidence="ECO:0000250"
FT   BINDING         568
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /evidence="ECO:0000250"
FT   BINDING         666
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /evidence="ECO:0000250"
FT   BINDING         670
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /evidence="ECO:0000250"
FT   MOD_RES         74
FT                   /note="N6-acetyllysine"
FT                   /evidence="ECO:0000269|PubMed:18723842,
FT                   ECO:0000269|PubMed:30274179"
FT   MUTAGEN         44
FT                   /note="M->A: In mutant 6A; abolished binding to lactate and
FT                   protein lactylation; when associated with A-70, A-171,
FT                   A-213 and 236-A--A-238."
FT                   /evidence="ECO:0000269|PubMed:38653238,
FT                   ECO:0000269|PubMed:39322678"
FT   MUTAGEN         70
FT                   /note="R->A: In mutant 6A; abolished binding to lactate and
FT                   protein lactylation; when associated with A-44, A-171,
FT                   A-213 and 236-A--A-238. In ATP-binding mutant; abolished
FT                   ability to mediate protein lactylation; when associated
FT                   with A-171 and A-240."
FT                   /evidence="ECO:0000269|PubMed:38653238,
FT                   ECO:0000269|PubMed:39322678"
FT   MUTAGEN         74
FT                   /note="K->A: Loss of alanylation activity on tRNA(Ala),
FT                   protein is not acetylated."
FT                   /evidence="ECO:0000269|PubMed:30274179"
FT   MUTAGEN         171
FT                   /note="W->A: In mutant 6A; abolished binding to lactate and
FT                   protein lactylation; when associated with A-44, A-70, A-213
FT                   and 236-A--A-238. In ATP-binding mutant; abolished ability
FT                   to mediate protein lactylation; when associated with A-70
FT                   and A-240."
FT                   /evidence="ECO:0000269|PubMed:38653238,
FT                   ECO:0000269|PubMed:39322678"
FT   MUTAGEN         179
FT                   /note="C->S: Inactivates the enzyme."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         182
FT                   /note="C->S: No inactivation."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         184
FT                   /note="E->Q: No inactivation."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         188
FT                   /note="D->N: No inactivation."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         189
FT                   /note="H->Q: Inactivates the enzyme."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         191
FT                   /note="D->N: No inactivation."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         192
FT                   /note="H->Q: Inactivates the enzyme."
FT                   /evidence="ECO:0000269|PubMed:1712632"
FT   MUTAGEN         213
FT                   /note="N->A: In mutant 6A; abolished binding to lactate and
FT                   protein lactylation; when associated with A-44, A-70, A-171
FT                   and 236-A--A-238."
FT                   /evidence="ECO:0000269|PubMed:38653238,
FT                   ECO:0000269|PubMed:39322678"
FT   MUTAGEN         236..238
FT                   /note="DTG->ATA: In mutant 6A; abolished binding to lactate
FT                   and protein lactylation; when associated with A-44, A-70,
FT                   A-171 and A-213."
FT                   /evidence="ECO:0000269|PubMed:38653238,
FT                   ECO:0000269|PubMed:39322678"
FT   MUTAGEN         236
FT                   /note="D->A,N: Decreases affinity for alanine without
FT                   improving discrimination against serine."
FT                   /evidence="ECO:0000269|PubMed:20010690"
FT   MUTAGEN         238
FT                   /note="G->A: Greatly decreases affinity for alanine with
FT                   only small changes in affinity for serine and glycine, in a
FT                   2-442 residue construct."
FT                   /evidence="ECO:0000269|PubMed:20010690"
FT   MUTAGEN         240
FT                   /note="G->A: In ATP-binding mutant; abolished ability to
FT                   mediate protein lactylation."
FT                   /evidence="ECO:0000269|PubMed:39322678"
FT   MUTAGEN         564
FT                   /note="H->A: No effect on tRNA editing."
FT                   /evidence="ECO:0000269|PubMed:12554667"
FT   MUTAGEN         567
FT                   /note="T->F: Complete loss of tRNA editing; when associated
FT                   with W-587 and F-666."
FT                   /evidence="ECO:0000269|PubMed:28362257"
FT   MUTAGEN         568
FT                   /note="H->A: No effect on tRNA editing."
FT                   /evidence="ECO:0000269|PubMed:12554667"
FT   MUTAGEN         584
FT                   /note="Q->H: Loss of mischarged tRNA editing activity; when
FT                   associated with A-666."
FT                   /evidence="ECO:0000269|PubMed:12554667,
FT                   ECO:0000269|PubMed:18723508"
FT   MUTAGEN         587
FT                   /note="S->W: Complete loss of tRNA editing; when associated
FT                   with F-567 and F-666."
FT                   /evidence="ECO:0000269|PubMed:28362257"
FT   MUTAGEN         666
FT                   /note="C->A: Loss of mischarged tRNA editing activity; when
FT                   associated with H-584 the effect is more pronounced."
FT                   /evidence="ECO:0000269|PubMed:12554667,
FT                   ECO:0000269|PubMed:18723508, ECO:0000269|PubMed:28362257"
FT   MUTAGEN         666
FT                   /note="C->F: Complete loss of tRNA editing; when associated
FT                   with F-567 and W-587."
FT                   /evidence="ECO:0000269|PubMed:28362257"
FT   MUTAGEN         670
FT                   /note="H->A: No effect on tRNA editing."
FT                   /evidence="ECO:0000269|PubMed:12554667"
FT   MUTAGEN         693
FT                   /note="R->K: Reduces specificity of editing activity for
FT                   tRNA(Ala), allows editing of tRNA(Thr)."
FT                   /evidence="ECO:0000269|PubMed:18172502"
FT   CONFLICT        29..33
FT                   /note="LVPHN -> RYPIT (in Ref. 4; no nucleotide entry and
FT                   5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        158
FT                   /note="G -> N (in Ref. 4; no nucleotide entry and 5;
FT                   AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        168
FT                   /note="D -> G (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        172
FT                   /note="Q -> R (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        175
FT                   /note="D -> G (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        180
FT                   /note="G -> D (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        584
FT                   /note="Q -> H (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        619
FT                   /note="Q -> L (in Ref. 5; AAA03208)"
FT                   /evidence="ECO:0000305"
FT   HELIX           5..18
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           25..28
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           43..47
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           48..52
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          60..69
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           78..80
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   TURN            81..83
FT                   /evidence="ECO:0007829|PDB:3HXU"
FT   STRAND          84..86
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          89..101
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           104..116
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   TURN            118..121
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           125..127
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          128..133
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           137..141
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           142..147
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           151..153
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          154..157
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          170..188
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   TURN            201..204
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          205..221
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          223..225
FT                   /evidence="ECO:0007829|PDB:3HY1"
FT   STRAND          227..240
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           241..248
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           254..256
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           258..271
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           279..296
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           305..324
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           331..334
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           335..342
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           343..346
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           347..351
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           353..368
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           371..383
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   STRAND          387..390
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           392..402
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           406..414
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   TURN            415..417
FT                   /evidence="ECO:0007829|PDB:3HY0"
FT   HELIX           422..440
FT                   /evidence="ECO:0007829|PDB:3HY0"
SQ   SEQUENCE   876 AA;  96032 MW;  73F69C69FCF8C08C CRC64;
     MSKSTAEIRQ AFLDFFHSKG HQVVASSSLV PHNDPTLLFT NAGMNQFKDV FLGLDKRNYS
     RATTSQRCVR AGGKHNDLEN VGYTARHHTF FEMLGNFSFG DYFKHDAIQF AWELLTSEKW
     FALPKERLWV TVYESDDEAY EIWEKEVGIP RERIIRIGDN KGAPYASDNF WQMGDTGPCG
     PCTEIFYDHG DHIWGGPPGS PEEDGDRYIE IWNIVFMQFN RQADGTMEPL PKPSVDTGMG
     LERIAAVLQH VNSNYDIDLF RTLIQAVAKV TGATDLSNKS LRVIADHIRS CAFLIADGVM
     PSNENRGYVL RRIIRRAVRH GNMLGAKETF FYKLVGPLID VMGSAGEDLK RQQAQVEQVL
     KTEEEQFART LERGLALLDE ELAKLSGDTL DGETAFRLYD TYGFPVDLTA DVCRERNIKV
     DEAGFEAAME EQRRRAREAS GFGADYNAMI RVDSASEFKG YDHLELNGKV TALFVDGKAV
     DAINAGQEAV VVLDQTPFYA ESGGQVGDKG ELKGANFSFA VEDTQKYGQA IGHIGKLAAG
     SLKVGDAVQA DVDEARRARI RLNHSATHLM HAALRQVLGT HVSQKGSLVN DKVLRFDFSH
     NEAMKPEEIR AVEDLVNTQI RRNLPIETNI MDLEAAKAKG AMALFGEKYD ERVRVLSMGD
     FSTELCGGTH ASRTGDIGLF RIISESGTAA GVRRIEAVTG EGAIATVHAD SDRLSEVAHL
     LKGDSNNLAD KVRSVLERTR QLEKELQQLK EQAAAQESAN LSSKAIDVNG VKLLVSELSG
     VEPKMLRTMV DDLKNQLGST IIVLATVVEG KVSLIAGVSK DVTDRVKAGE LIGMVAQQVG
     GKGGGRPDMA QAGGTDAAAL PAALASVKGW VSAKLQ
//