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Protein

Isoleucine--tRNA ligase 2

Gene

ileS2

Organism
Pseudomonas fluorescens
Status
Reviewed-Annotation score: Annotation score: 4 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the attachment of isoleucine to tRNA(Ile). As IleRS can inadvertently accommodate and process structurally similar amino acids such as valine, to avoid such errors it has two additional distinct tRNA(Ile)-dependent editing activities. One activity is designated as 'pretransfer' editing and involves the hydrolysis of activated Val-AMP. The other activity is designated 'posttransfer' editing and involves deacylation of mischarged Val-tRNA(Ile) (By similarity).By similarity
Confers high-level resistance to the antibiotic mupirocin (pseudomonic acid A), an Ile-analog produced by P.fluorescens NCIMB 10586 itself that competitively inhibits activation by Ile-tRNA synthetase, thus inhibiting protein biosynthesis.

Catalytic activityi

ATP + L-isoleucine + tRNA(Ile) = AMP + diphosphate + L-isoleucyl-tRNA(Ile).

Cofactori

Zn2+By similarity

Kineticsi

  1. KM=10 µM for isoleucine
  2. KM=330 µM for ATP

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei592 – 5921ATPBy similarity

    GO - Molecular functioni

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Aminoacyl-tRNA synthetase, Ligase

    Keywords - Biological processi

    Antibiotic resistance, Protein biosynthesis

    Keywords - Ligandi

    ATP-binding, Metal-binding, Nucleotide-binding, Zinc

    Enzyme and pathway databases

    BRENDAi6.1.1.5. 5121.
    SABIO-RKQ8L1B1.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Isoleucine--tRNA ligase 2 (EC:6.1.1.5)
    Alternative name(s):
    Isoleucyl-tRNA synthetase 2
    Short name:
    IleRS 2
    Gene namesi
    Name:ileS2
    Synonyms:mupM
    OrganismiPseudomonas fluorescens
    Taxonomic identifieri294 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaPseudomonadalesPseudomonadaceaePseudomonas

    Subcellular locationi

    GO - Cellular componenti

    Complete GO annotation...

    Keywords - Cellular componenti

    Cytoplasm

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Initiator methioninei1 – 11Removed1 Publication
    Chaini2 – 10301029Isoleucine--tRNA ligase 2PRO_0000098556Add
    BLAST

    Interactioni

    Subunit structurei

    Monomer.By similarity

    Structurei

    3D structure databases

    ProteinModelPortaliQ8L1B1.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Motif

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Motifi48 – 5811"HIGH" regionAdd
    BLAST
    Motifi589 – 5935"KMSKS" region

    Domaini

    IleRS has two distinct active sites: one for aminoacylation and one for editing. The misactivated valine is translocated from the active site to the editing site, which sterically excludes the correctly activated isoleucine. The single editing site contains two valyl binding pockets, one specific for each substrate (Val-AMP or Val-tRNA(Ile)) (By similarity).By similarity

    Sequence similaritiesi

    Family and domain databases

    Gene3Di1.10.730.10. 1 hit.
    3.40.50.620. 2 hits.
    3.90.740.10. 1 hit.
    HAMAPiMF_02003. Ile_tRNA_synth_type2.
    InterProiIPR001412. aa-tRNA-synth_I_CS.
    IPR002300. aa-tRNA-synth_Ia.
    IPR002301. Ile-tRNA-ligase.
    IPR023586. Ile-tRNA-ligase_type2.
    IPR013155. M/V/L/I-tRNA-synth_anticd-bd.
    IPR014729. Rossmann-like_a/b/a_fold.
    IPR009080. tRNAsynth_1a_anticodon-bd.
    IPR009008. Val/Leu/Ile-tRNA-synth_edit.
    [Graphical view]
    PfamiPF08264. Anticodon_1. 1 hit.
    PF00133. tRNA-synt_1. 1 hit.
    [Graphical view]
    PRINTSiPR00984. TRNASYNTHILE.
    SUPFAMiSSF47323. SSF47323. 1 hit.
    SSF50677. SSF50677. 1 hit.
    TIGRFAMsiTIGR00392. ileS. 1 hit.
    PROSITEiPS00178. AA_TRNA_LIGASE_I. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    Q8L1B1-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MSTEGSGPVR FPAMEDAVLE RWEKEKTFEQ SISAREGKPV YVFYDGPPFA
    60 70 80 90 100
    TGLPHYGHIL TSYIKDVIPR YQTMLGKQVP RRWGWDCHGL PVEFEVEKAM
    110 120 130 140 150
    GFKSKRDILE FGVEQFNDEC RELVLKYADD WRGFVNRMGR WVDFDGAYKT
    160 170 180 190 200
    MDNDYMESVL WGFKTLHDKG HVYEGGKIVP YCVRCQTVLS NFEARLDDAF
    210 220 230 240 250
    RPRRDMSAYV KFRQQDRPDT FFLAWTTTPW TLPANVALAV AADENYVCIE
    260 270 280 290 300
    HGEERLWLAE GCLGGLFDEP VILERCTGAE LAGLRYLPVV GEVIDASAHR
    310 320 330 340 350
    VVTADFVQMG DGSGIVHIAP AFGEDDALLG QQYELPAPNP VRDDGTFSDA
    360 370 380 390 400
    VAQYAGQNIF EATPRILADL KSSGLLFKQE QIEHNYPHCW RCDNPLIYRA
    410 420 430 440 450
    VESWFIRASA LREQLVENNS QVNWVPEHVK EGRFGDWIRN ARDWAVSRNR
    460 470 480 490 500
    FWGAPIPVWR CDQCGTVEVM GSIAQIEARS GRKVEDLHVP HIDEHRFACQ
    510 520 530 540 550
    CCEGTMSRVT GVFDCWFESG AMPFASRHYP FENKQEFEQT FPADFIVEYL
    560 570 580 590 600
    AQTRGWFYTM MVISTGCFEQ NPFKNAMCHG VILAKDGRKM SKRLKNYPNP
    610 620 630 640 650
    MDLMQTHGSD ALRVALLASP VCKGEDIKFS EESVRDVVRR YHLLFWNCLQ
    660 670 680 690 700
    FYKTFTEIDQ FSPSGDLGQP LDNVLDHYLL HELAALESDI KMWMESLDFS
    710 720 730 740 750
    KIYSRIEVFI NVLSTWYLRL NKARIWRDGL DDDKRQCYEV LHYALSNFAR
    760 770 780 790 800
    LLAPFMPFLA EAVYTELGYA DSVHLQDWPS IDRQYLSYEL ADEMSSLRNL
    810 820 830 840 850
    IASVRNVRET NGVSQKFPLR SIRVAGIEQA VLERYAQFLE EELNVKQVQW
    860 870 880 890 900
    AADADEWAQP VVVLIFSLLG KRLGPAMKAV TTAVKAGEYV IDEQGGLVAA
    910 920 930 940 950
    GQTIQPHEFE RRLTVRDTLN NVGIVENMVV WLDLDIDASL KREGAVRELN
    960 970 980 990 1000
    RRLQDLRKKA KLGYTEKVDI AVLGGAYVDE ILVHHEDWLK SQLLVQSLLR
    1010 1020 1030
    SDLEAPLAVD EVELPEGDPV RIQLRRSVLA
    Length:1,030
    Mass (Da):117,875
    Last modified:January 23, 2007 - v3
    Checksum:i2D930693BE1DC935
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti175 – 1751G → R in AAL85500 (PubMed:12652905).Curated
    Sequence conflicti400 – 4001A → T in AAM12927 (Ref. 2) Curated
    Sequence conflicti667 – 6671L → P in AAM12927 (Ref. 2) Curated
    Sequence conflicti671 – 6711L → P in AAM12927 (Ref. 2) Curated
    Sequence conflicti717 – 7171Y → H in AAM12927 (Ref. 2) Curated
    Sequence conflicti805 – 8051R → C in AAM12927 (Ref. 2) Curated
    Sequence conflicti886 – 8861A → V in AAM12927 (Ref. 2) Curated
    Sequence conflicti939 – 9391S → P in AAM12927 (Ref. 2) Curated

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AY079084 Genomic DNA. Translation: AAL85500.1.
    AF318063 Genomic DNA. Translation: AAM12927.1.
    AB062785 Genomic DNA. Translation: BAC07171.1.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AY079084 Genomic DNA. Translation: AAL85500.1.
    AF318063 Genomic DNA. Translation: AAM12927.1.
    AB062785 Genomic DNA. Translation: BAC07171.1.

    3D structure databases

    ProteinModelPortaliQ8L1B1.
    ModBaseiSearch...
    MobiDBiSearch...

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Enzyme and pathway databases

    BRENDAi6.1.1.5. 5121.
    SABIO-RKQ8L1B1.

    Family and domain databases

    Gene3Di1.10.730.10. 1 hit.
    3.40.50.620. 2 hits.
    3.90.740.10. 1 hit.
    HAMAPiMF_02003. Ile_tRNA_synth_type2.
    InterProiIPR001412. aa-tRNA-synth_I_CS.
    IPR002300. aa-tRNA-synth_Ia.
    IPR002301. Ile-tRNA-ligase.
    IPR023586. Ile-tRNA-ligase_type2.
    IPR013155. M/V/L/I-tRNA-synth_anticd-bd.
    IPR014729. Rossmann-like_a/b/a_fold.
    IPR009080. tRNAsynth_1a_anticodon-bd.
    IPR009008. Val/Leu/Ile-tRNA-synth_edit.
    [Graphical view]
    PfamiPF08264. Anticodon_1. 1 hit.
    PF00133. tRNA-synt_1. 1 hit.
    [Graphical view]
    PRINTSiPR00984. TRNASYNTHILE.
    SUPFAMiSSF47323. SSF47323. 1 hit.
    SSF50677. SSF50677. 1 hit.
    TIGRFAMsiTIGR00392. ileS. 1 hit.
    PROSITEiPS00178. AA_TRNA_LIGASE_I. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    1. "Analysis of rILERS, an isoleucyl-tRNA synthetase gene associated with mupirocin production by Pseudomonas fluorescens NCIMB 10586."
      Rangaswamy V., Hernandez-Guzman G., Shufran K.A., Bender C.L.
      DNA Seq. 13:343-351(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: ATCC 49323 / NCIMB 10586.
    2. "Nucleotide sequence of a 75 kb region of the chromosome of Pseudomonas fluorescens NCIMB 10586 required for biosynthesis of the polyketide antibiotic mupirocin."
      El-Sayed A.K., Hothersall J., Cooper S.M., Stephens E., Thomas C.M.
      Submitted (JAN-2002) to the EMBL/GenBank/DDBJ databases
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: ATCC 49323 / NCIMB 10586.
    3. "How does Pseudomonas fluorescens avoid suicide from its antibiotic pseudomonic acid? Evidence for two evolutionarily distinct isoleucyl-tRNA synthetases conferring self-defense."
      Yanagisawa T., Kawakami M.
      J. Biol. Chem. 278:25887-25894(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-13, RESISTANCE TO MUPIROCIN.
      Strain: ATCC 49323 / NCIMB 10586.

    Entry informationi

    Entry nameiSYI2_PSEFL
    AccessioniPrimary (citable) accession number: Q8L1B1
    Secondary accession number(s): Q8GM72, Q8RL58
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: December 6, 2005
    Last sequence update: January 23, 2007
    Last modified: January 7, 2015
    This is version 66 of the entry and version 3 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    P.fluorescens NCIMB 10586 possesses two distinct IleRSs (IleRS-R1 and IleRS-R2), each with a different level of sensitivity to mupirocin. Purified IleRs-R2 shows no sensitivity to mupirocin even at a concentration of 5 mM, 100'000 fold higher than the Ki value of IleRS-R1.

    Keywords - Technical termi

    Direct protein sequencing

    Documents

    1. Aminoacyl-tRNA synthetases
      List of aminoacyl-tRNA synthetase entries
    2. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.