SYP_ECOLI
ID SYP_ECOLI Reviewed; 572 AA.
AC P16659; P78272; Q59430;
DT 01-AUG-1990, integrated into UniProtKB/Swiss-Prot.
DT 29-AUG-2003, sequence version 4.
DT 03-AUG-2022, entry version 191.
DE RecName: Full=Proline--tRNA ligase;
DE EC=6.1.1.15;
DE AltName: Full=Global RNA synthesis factor;
DE AltName: Full=Prolyl-tRNA synthetase;
DE Short=ProRS;
GN Name=proS; Synonyms=drpA; OrderedLocusNames=b0194, JW0190;
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 1-12.
RX PubMed=2203971; DOI=10.1038/347203a0;
RA Eriani G., Delarue M., Poch O., Gangloff J., Moras D.;
RT "Partition of tRNA synthetases into two classes based on mutually exclusive
RT sets of sequence motifs.";
RL Nature 347:203-206(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=1688424; DOI=10.1128/jb.172.1.281-286.1990;
RA Zhou Z., Syvanen M.;
RT "Identification and sequence of the drpA gene from Escherichia coli.";
RL J. Bacteriol. 172:281-286(1990).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RA Takemoto K., Mori H., Murayama N., Kataoka K., Yano M., Itoh T.,
RA Yamamoto Y., Inokuchi H., Miki T., Hatada E., Fukuda R., Ichihara S.,
RA Mizuno T., Makino K., Nakata A., Yura T., Sampei G., Mizobuchi K.;
RT "Systematic sequencing of the Escherichia coli genome: analysis of the 4.0
RT - 6.0 min (189,987 - 281,416bp) region.";
RL Submitted (FEB-1996) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [5]
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 [6]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA], AND SEQUENCE REVISION TO
RP 26-27; 205 AND 568-572.
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 [7]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-155.
RC STRAIN=K12;
RA Miyamoto K.;
RL Submitted (APR-1993) to the EMBL/GenBank/DDBJ databases.
RN [8]
RP PROTEIN SEQUENCE OF 1-12.
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 [9]
RP EDITING ACTIVITY, KINETIC PARAMETERS, AND MUTAGENESIS OF CYS-443.
RX PubMed=10922054; DOI=10.1073/pnas.97.16.8916;
RA Beuning P.J., Musier-Forsyth K.;
RT "Hydrolytic editing by a class II aminoacyl-tRNA synthetase.";
RL Proc. Natl. Acad. Sci. U.S.A. 97:8916-8920(2000).
RN [10]
RP EDITING ACTIVITY, AND KINETIC PARAMETERS.
RX PubMed=11408489; DOI=10.1074/jbc.m104761200;
RA Beuning P.J., Musier-Forsyth K.;
RT "Species-specific differences in amino acid editing by class II prolyl-tRNA
RT synthetase.";
RL J. Biol. Chem. 276:30779-30785(2001).
RN [11]
RP MUTAGENESIS OF THR-257; LYS-279; ASP-350; HIS-369; ASP-378; ASP-386 AND
RP ASP-394.
RX PubMed=12033945; DOI=10.1021/bi012178j;
RA Wong F.-C., Beuning P.J., Nagan M., Shiba K., Musier-Forsyth K.;
RT "Functional role of the prokaryotic proline-tRNA synthetase insertion
RT domain in amino acid editing.";
RL Biochemistry 41:7108-7115(2002).
RN [12]
RP PROLINE AND CYSTEINE ACTIVATION, EDITING ACTIVITY, AND KINETIC PARAMETERS.
RX PubMed=12130657; DOI=10.1074/jbc.m206928200;
RA Ahel I., Stathopoulos C., Ambrogelly A., Sauerwald A., Toogood H.,
RA Hartsch T., Soell D.;
RT "Cysteine activation is an inherent in vitro property of prolyl-tRNA
RT synthetases.";
RL J. Biol. Chem. 277:34743-34748(2002).
RN [13]
RP EDITING DOMAIN.
RX PubMed=14530268; DOI=10.1074/jbc.m309627200;
RA Wong F.-C., Beuning P.J., Silvers C., Musier-Forsyth K.;
RT "An isolated class II aminoacyl-tRNA synthetase insertion domain is
RT functional in amino acid editing.";
RL J. Biol. Chem. 278:52857-52864(2003).
CC -!- FUNCTION: Catalyzes the attachment of proline to tRNA(Pro) in a two-
CC step reaction: proline is first activated by ATP to form Pro-AMP and
CC then transferred to the acceptor end of tRNA(Pro). As ProRS can
CC inadvertently accommodate and process non-cognate amino acids such as
CC alanine and cysteine, to avoid such errors it has two additional
CC distinct editing activities against alanine. One activity is designated
CC as 'pretransfer' editing and involves the tRNA(Pro)-independent
CC hydrolysis of activated Ala-AMP. The other activity is designated
CC 'posttransfer' editing and involves deacylation of mischarged Ala-
CC tRNA(Pro). Misacylated Cys-tRNA(Pro) is not edited by ProRS, but
CC instead may be edited in trans by YbaK.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-proline + tRNA(Pro) = AMP + diphosphate + L-prolyl-
CC tRNA(Pro); Xref=Rhea:RHEA:14305, Rhea:RHEA-COMP:9700, Rhea:RHEA-
CC COMP:9702, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:60039,
CC ChEBI:CHEBI:78442, ChEBI:CHEBI:78532, ChEBI:CHEBI:456215;
CC EC=6.1.1.15;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.25 mM for proline {ECO:0000269|PubMed:10922054,
CC ECO:0000269|PubMed:11408489, ECO:0000269|PubMed:12130657};
CC KM=140 mM for alanine {ECO:0000269|PubMed:10922054,
CC ECO:0000269|PubMed:11408489, ECO:0000269|PubMed:12130657};
CC KM=0.17 mM for cysteine {ECO:0000269|PubMed:10922054,
CC ECO:0000269|PubMed:11408489, ECO:0000269|PubMed:12130657};
CC -!- SUBUNIT: Homodimer.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- DOMAIN: Consists of three domains: the N-terminal catalytic domain, the
CC editing domain and the C-terminal anticodon-binding domain.
CC -!- SIMILARITY: Belongs to the class-II aminoacyl-tRNA synthetase family.
CC ProS type 1 subfamily. {ECO:0000305}.
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DR EMBL; X55518; CAA39134.1; -; Genomic_DNA.
DR EMBL; M97858; AAA24420.1; -; Genomic_DNA.
DR EMBL; M32357; AAA23710.1; -; Genomic_DNA.
DR EMBL; U70214; AAB08622.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73305.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA77870.2; -; Genomic_DNA.
DR EMBL; D15061; BAA03654.1; -; Genomic_DNA.
DR PIR; B64744; YPEC.
DR RefSeq; NP_414736.1; NC_000913.3.
DR RefSeq; WP_001260717.1; NZ_LN832404.1.
DR AlphaFoldDB; P16659; -.
DR SMR; P16659; -.
DR BioGRID; 4261629; 32.
DR DIP; DIP-10573N; -.
DR IntAct; P16659; 20.
DR STRING; 511145.b0194; -.
DR CarbonylDB; P16659; -.
DR SWISS-2DPAGE; P16659; -.
DR jPOST; P16659; -.
DR PaxDb; P16659; -.
DR PRIDE; P16659; -.
DR EnsemblBacteria; AAC73305; AAC73305; b0194.
DR EnsemblBacteria; BAA77870; BAA77870; BAA77870.
DR GeneID; 949116; -.
DR KEGG; ecj:JW0190; -.
DR KEGG; eco:b0194; -.
DR PATRIC; fig|1411691.4.peg.2084; -.
DR EchoBASE; EB0763; -.
DR eggNOG; COG0442; Bacteria.
DR HOGENOM; CLU_016739_0_0_6; -.
DR InParanoid; P16659; -.
DR OMA; NCDYAAN; -.
DR PhylomeDB; P16659; -.
DR BioCyc; EcoCyc:PROS-MON; -.
DR BioCyc; MetaCyc:PROS-MON; -.
DR BRENDA; 6.1.1.15; 2026.
DR SABIO-RK; P16659; -.
DR PRO; PR:P16659; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0043906; F:Ala-tRNA(Pro) hydrolase activity; IDA:UniProtKB.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0004827; F:proline-tRNA ligase activity; IDA:EcoCyc.
DR GO; GO:0006433; P:prolyl-tRNA aminoacylation; IMP:EcoCyc.
DR CDD; cd00861; ProRS_anticodon_short; 1.
DR CDD; cd00779; ProRS_core_prok; 1.
DR Gene3D; 3.30.930.10; -; 2.
DR Gene3D; 3.40.50.800; -; 1.
DR Gene3D; 3.90.960.10; -; 1.
DR HAMAP; MF_01569; Pro_tRNA_synth_type1; 1.
DR InterPro; IPR002314; aa-tRNA-synt_IIb.
DR InterPro; IPR006195; aa-tRNA-synth_II.
DR InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL.
DR InterPro; IPR004154; Anticodon-bd.
DR InterPro; IPR036621; Anticodon-bd_dom_sf.
DR InterPro; IPR002316; Pro-tRNA-ligase_IIa.
DR InterPro; IPR004500; Pro-tRNA-synth_IIa_bac-type.
DR InterPro; IPR023717; Pro-tRNA-Synthase_IIa_type1.
DR InterPro; IPR044140; ProRS_anticodon_short.
DR InterPro; IPR033730; ProRS_core_prok.
DR InterPro; IPR036754; YbaK/aa-tRNA-synt-asso_dom_sf.
DR InterPro; IPR007214; YbaK/aa-tRNA-synth-assoc-dom.
DR Pfam; PF03129; HGTP_anticodon; 1.
DR Pfam; PF00587; tRNA-synt_2b; 1.
DR Pfam; PF04073; tRNA_edit; 1.
DR PIRSF; PIRSF001535; ProRS_1; 1.
DR PRINTS; PR01046; TRNASYNTHPRO.
DR SUPFAM; SSF55681; SSF55681; 1.
DR SUPFAM; SSF55826; SSF55826; 1.
DR TIGRFAMs; TIGR00409; proS_fam_II; 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 CHAIN 1..572
FT /note="Proline--tRNA ligase"
FT /id="PRO_0000139329"
FT MUTAGEN 257
FT /note="T->A: Reduces the posttransfer editing activity 5-
FT fold, with little change in both the aminoacylation and the
FT pretransfer editing activities."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 279
FT /note="K->A: Severely affects the posttransfer editing
FT activity, with little change in both the aminoacylation and
FT the pretransfer editing activities."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 350
FT /note="D->A: Abolishes the pretransfer editing activity and
FT reduces the aminoacylation activity 20-fold and the
FT posttransfer editing activity 5-fold."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 369
FT /note="H->A: Reduces both the aminoacylation and the
FT pretransfer editing activities 2.5-fold, and the
FT posttransfer editing activity 5-fold. Loss of specificity
FT in deacylation."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 378
FT /note="D->A: Little change in both the aminoacylation and
FT the editing activities."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 386
FT /note="D->A: Reduces the posttransfer editing activity
FT approximately 2-fold, with little change in both the
FT aminoacylation and the pretransfer editing activities."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 394
FT /note="D->A: Reduces the posttransfer editing activity
FT approximately 2-fold, with little change in both the
FT aminoacylation and the pretransfer editing activities."
FT /evidence="ECO:0000269|PubMed:12033945"
FT MUTAGEN 443
FT /note="C->G: 150-fold decrease in posttransfer editing
FT activity against alanine, without change neither in
FT pretransfer editing nor in aminoacylation."
FT /evidence="ECO:0000269|PubMed:10922054"
FT CONFLICT 26..27
FT /note="ML -> IV (in Ref. 1)"
FT /evidence="ECO:0000305"
FT CONFLICT 127..128
FT /note="QL -> HV (in Ref. 2; AAA23710)"
FT /evidence="ECO:0000305"
FT CONFLICT 205..216
FT /note="SASHEFQVLAQS -> RPLTNSRCWRR (in Ref. 2; AAA23710)"
FT /evidence="ECO:0000305"
FT CONFLICT 205
FT /note="S -> Q (in Ref. 1)"
FT /evidence="ECO:0000305"
FT CONFLICT 516..517
FT /note="PG -> RA (in Ref. 2)"
FT /evidence="ECO:0000305"
FT CONFLICT 518..572
FT /note="Missing (in Ref. 2)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 572 AA; 63693 MW; 312D29390E91C486 CRC64;
MRTSQYLLST LKETPADAEV ISHQLMLRAG MIRKLASGLY TWLPTGVRVL KKVENIVREE
MNNAGAIEVS MPVVQPADLW QESGRWEQYG PELLRFVDRG ERPFVLGPTH EEVITDLIRN
ELSSYKQLPL NFYQIQTKFR DEVRPRFGVM RSREFLMKDA YSFHTSQESL QETYDAMYAA
YSKIFSRMGL DFRAVQADTG SIGGSASHEF QVLAQSGEDD VVFSDTSDYA ANIELAEAIA
PKEPRAAATQ EMTLVDTPNA KTIAELVEQF NLPIEKTVKT LLVKAVEGSS FPQVALLVRG
DHELNEVKAE KLPQVASPLT FATEEEIRAV VKAGPGSLGP VNMPIPVVID RTVAAMSDFA
AGANIDGKHY FGINWDRDVA TPEVADIRNV VAGDPSPDGQ GRLLIKRGIE VGHIFQLGTK
YSEALKASVQ GEDGRNQILT MGCYGIGVTR VVAAAIEQNY DERGIVWPDA IAPFQVAILP
MNMHKSFRVQ ELAEKLYSEL RAQGIEVLLD DRKERPGVMF ADMELIGIPH TIVLGDRNLD
NDDIEYKYRR NGEKQLIKTG DIVEYLVKQI KG
