Skip Header

You are using a version of browser that may not display all the features of this website. Please consider upgrading your browser.
Protein

Asparagine synthetase B [glutamine-hydrolyzing]

Gene

asnB

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the ATP-dependent conversion of aspartate into asparagine, using glutamine as a source of nitrogen. Can also use ammonia as the nitrogen source in vitro, albeit with lower efficiency. As nucleotide substrates, ATP and dATP are utilized at a similar rate in both the glutamine- and ammonia-dependent reactions, whereas GTP utilization is only 15% that of ATP, and CTP, UTP, ITP and XTP are very poor or not substrates. Also exhibits glutaminase activity.3 Publications

Catalytic activityi

ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate.2 Publications

Enzyme regulationi

Glutamine-dependent asparagine synthesis activity can be inhibited by aspartic acid analogs (such as a sulfinate derivative and (2S,3R)-2-amino-3-methylsuccinate) in vitro; the inhibition is competitive with respect to aspartate.1 Publication

Kineticsi

  1. KM=17 mM for ammonia2 Publications
  2. KM=0.53 mM for aspartate (when assaying the ammonia-dependent synthetase reaction)2 Publications
  3. KM=0.85 mM for aspartate (when assaying the glutamine-dependent synthetase reaction)2 Publications
  4. KM=0.26 mM for ATP (when assaying the glutamine-dependent synthetase reaction)2 Publications
  5. KM=0.66 mM for glutamine (when assaying the glutamine-dependent synthetase reaction)2 Publications

    pH dependencei

    Optimum pH is 6.5-8.1 Publication

    Pathwayi

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Active sitei2 – 21For GATase activity
    Binding sitei99 – 991Glutamine
    Binding sitei233 – 2331ATP; via carbonyl oxygen
    Binding sitei273 – 2731ATP; via amide nitrogen and carbonyl oxygen
    Sitei349 – 3491Important for beta-aspartyl-AMP intermediate formation

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi347 – 3482ATP

    GO - Molecular functioni

    • amino acid binding Source: UniProtKB
    • asparagine synthase (glutamine-hydrolyzing) activity Source: UniProtKB
    • aspartate-ammonia ligase activity Source: UniProtKB
    • ATP binding Source: UniProtKB
    • identical protein binding Source: IntAct

    GO - Biological processi

    • asparagine biosynthetic process Source: UniProtKB
    • cellular amino acid biosynthetic process Source: EcoliWiki
    • cellular amino acid catabolic process Source: EcoliWiki
    • glutamine metabolic process Source: EcoliWiki
    • L-asparagine biosynthetic process Source: UniProtKB-UniPathway
    Complete GO annotation...

    Keywords - Molecular functioni

    Ligase

    Keywords - Biological processi

    Amino-acid biosynthesis, Asparagine biosynthesis

    Keywords - Ligandi

    ATP-binding, Nucleotide-binding

    Enzyme and pathway databases

    BioCyciEcoCyc:ASNSYNB-MONOMER.
    ECOL316407:JW0660-MONOMER.
    MetaCyc:ASNSYNB-MONOMER.
    BRENDAi6.3.5.4. 2026.
    UniPathwayiUPA00134; UER00195.

    Protein family/group databases

    MEROPSiC44.976.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Asparagine synthetase B [glutamine-hydrolyzing] (EC:6.3.5.4)
    Short name:
    AS-B
    Gene namesi
    Name:asnB
    Ordered Locus Names:b0674, JW0660
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG10092. asnB.

    Subcellular locationi

    GO - Cellular componenti

    • cytoplasm Source: EcoliWiki
    Complete GO annotation...

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi2 – 21C → A or S: Loss of glutamine-dependent activity but no effect on ammonia-dependent asparagine synthetase activity. 1 Publication
    Mutagenesisi30 – 301H → A: 4,5-fold decrease in glutamine affinity. 1 Publication
    Mutagenesisi34 – 341D → N or E: Little effect on the kinetic properties. 1 Publication
    Mutagenesisi81 – 811H → A: 5-fold decrease in glutamine affinity. 1 Publication
    Mutagenesisi105 – 1051A → H: Little effect on the kinetic properties. 1 Publication
    Mutagenesisi349 – 3491E → A or Q: Loss of glutamine- and ammonia-dependent synthetase activity, but still exhibits glutaminase activity. 1 Publication
    Mutagenesisi349 – 3491E → D: 5-fold increase in affinity for asparate when assaying both the glutamine- and ammonia-dependent synthetase reactions, and 2-fold decrease in kcat for these reactions. Modifies the product glutamate/asparagine stoichiometry. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Initiator methioninei1 – 11Removed1 Publication
    Chaini2 – 554553Asparagine synthetase B [glutamine-hydrolyzing]PRO_0000056931Add
    BLAST

    Proteomic databases

    PaxDbiP22106.
    PRIDEiP22106.

    Expressioni

    Gene expression databases

    GenevestigatoriP22106.

    Interactioni

    Subunit structurei

    Homodimer.1 Publication

    Binary interactionsi

    WithEntry#Exp.IntActNotes
    itself3EBI-549123,EBI-549123

    Protein-protein interaction databases

    DIPiDIP-9177N.
    IntActiP22106. 7 interactions.
    MINTiMINT-1306603.
    STRINGi511145.b0674.

    Structurei

    Secondary structure

    1
    554
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi3 – 97Combined sources
    Helixi14 – 2613Combined sources
    Helixi27 – 315Combined sources
    Beta strandi35 – 406Combined sources
    Beta strandi42 – 509Combined sources
    Turni56 – 583Combined sources
    Beta strandi61 – 644Combined sources
    Beta strandi70 – 789Combined sources
    Helixi81 – 888Combined sources
    Turni89 – 913Combined sources
    Helixi99 – 1013Combined sources
    Helixi102 – 1109Combined sources
    Turni111 – 1144Combined sources
    Helixi115 – 1173Combined sources
    Beta strandi120 – 1289Combined sources
    Turni129 – 1324Combined sources
    Beta strandi133 – 1386Combined sources
    Beta strandi147 – 1504Combined sources
    Beta strandi156 – 1616Combined sources
    Helixi162 – 1643Combined sources
    Turni165 – 1684Combined sources
    Beta strandi170 – 1745Combined sources
    Beta strandi179 – 1824Combined sources
    Turni183 – 1853Combined sources
    Beta strandi187 – 1904Combined sources
    Helixi195 – 1973Combined sources
    Helixi199 – 2024Combined sources
    Helixi209 – 22315Combined sources
    Beta strandi230 – 2334Combined sources
    Helixi238 – 25013Combined sources
    Beta strandi269 – 2757Combined sources
    Helixi279 – 29113Combined sources
    Beta strandi294 – 2996Combined sources
    Helixi302 – 31615Combined sources
    Helixi321 – 33919Combined sources
    Beta strandi344 – 3463Combined sources
    Helixi351 – 3555Combined sources
    Helixi359 – 3635Combined sources
    Helixi367 – 38014Combined sources
    Helixi381 – 3833Combined sources
    Helixi385 – 39410Combined sources
    Turni395 – 3973Combined sources
    Beta strandi399 – 4013Combined sources
    Helixi403 – 4053Combined sources
    Helixi407 – 4159Combined sources
    Helixi418 – 4203Combined sources
    Helixi431 – 4377Combined sources
    Helixi438 – 4403Combined sources
    Helixi443 – 4464Combined sources
    Helixi460 – 47213Combined sources
    Helixi475 – 4795Combined sources
    Helixi481 – 4844Combined sources
    Helixi493 – 50513Combined sources
    Helixi509 – 5146Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1CT9X-ray2.00A/B/C/D2-554[»]
    ProteinModelPortaliP22106.
    SMRiP22106. Positions 3-517.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP22106.

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini2 – 186185Glutamine amidotransferase type-2PROSITE-ProRule annotationAdd
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni50 – 545Glutamine binding
    Regioni75 – 773Glutamine binding

    Sequence similaritiesi

    Belongs to the asparagine synthetase family.Curated
    Contains 1 glutamine amidotransferase type-2 domain.PROSITE-ProRule annotation

    Keywords - Domaini

    Glutamine amidotransferase

    Phylogenomic databases

    eggNOGiCOG0367.
    HOGENOMiHOG000027493.
    InParanoidiP22106.
    KOiK01953.
    OMAiDWSGIYS.
    OrthoDBiEOG6J48H7.
    PhylomeDBiP22106.

    Family and domain databases

    Gene3Di3.40.50.620. 1 hit.
    3.60.20.10. 1 hit.
    InterProiIPR006426. Asn_synth_AEB.
    IPR001962. Asn_synthase.
    IPR017932. GATase_2_dom.
    IPR000583. GATase_dom.
    IPR029055. Ntn_hydrolases_N.
    IPR014729. Rossmann-like_a/b/a_fold.
    [Graphical view]
    PfamiPF00733. Asn_synthase. 1 hit.
    PF13537. GATase_7. 1 hit.
    [Graphical view]
    SUPFAMiSSF56235. SSF56235. 1 hit.
    TIGRFAMsiTIGR01536. asn_synth_AEB. 1 hit.
    PROSITEiPS51278. GATASE_TYPE_2. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

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

    P22106-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MCSIFGVFDI KTDAVELRKK ALELSRLMRH RGPDWSGIYA SDNAILAHER
    60 70 80 90 100
    LSIVDVNAGA QPLYNQQKTH VLAVNGEIYN HQALRAEYGD RYQFQTGSDC
    110 120 130 140 150
    EVILALYQEK GPEFLDDLQG MFAFALYDSE KDAYLIGRDH LGIIPLYMGY
    160 170 180 190 200
    DEHGQLYVAS EMKALVPVCR TIKEFPAGSY LWSQDGEIRS YYHRDWFDYD
    210 220 230 240 250
    AVKDNVTDKN ELRQALEDSV KSHLMSDVPY GVLLSGGLDS SIISAITKKY
    260 270 280 290 300
    AARRVEDQER SEAWWPQLHS FAVGLPGSPD LKAAQEVANH LGTVHHEIHF
    310 320 330 340 350
    TVQEGLDAIR DVIYHIETYD VTTIRASTPM YLMSRKIKAM GIKMVLSGEG
    360 370 380 390 400
    SDEVFGGYLY FHKAPNAKEL HEETVRKLLA LHMYDCARAN KAMSAWGVEA
    410 420 430 440 450
    RVPFLDKKFL DVAMRINPQD KMCGNGKMEK HILRECFEAY LPASVAWRQK
    460 470 480 490 500
    EQFSDGVGYS WIDTLKEVAA QQVSDQQLET ARFRFPYNTP TSKEAYLYRE
    510 520 530 540 550
    IFEELFPLPS AAECVPGGPS VACSSAKAIE WDEAFKKMDD PSGRAVGVHQ

    SAYK
    Length:554
    Mass (Da):62,659
    Last modified:January 23, 2007 - v3
    Checksum:i9091B269112C9F5C
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    J05554 Genomic DNA. Translation: AAA23498.1.
    U00096 Genomic DNA. Translation: AAC73768.1.
    AP009048 Genomic DNA. Translation: BAA35317.1.
    PIRiA36616. AJECN.
    RefSeqiNP_415200.1. NC_000913.3.

    Genome annotation databases

    EnsemblBacteriaiAAC73768; AAC73768; b0674.
    BAA35317; BAA35317; BAA35317.
    GeneIDi945281.
    KEGGiecj:Y75_p0653.
    eco:b0674.
    PATRICi32116529. VBIEscCol129921_0697.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    J05554 Genomic DNA. Translation: AAA23498.1.
    U00096 Genomic DNA. Translation: AAC73768.1.
    AP009048 Genomic DNA. Translation: BAA35317.1.
    PIRiA36616. AJECN.
    RefSeqiNP_415200.1. NC_000913.3.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1CT9X-ray2.00A/B/C/D2-554[»]
    ProteinModelPortaliP22106.
    SMRiP22106. Positions 3-517.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-9177N.
    IntActiP22106. 7 interactions.
    MINTiMINT-1306603.
    STRINGi511145.b0674.

    Protein family/group databases

    MEROPSiC44.976.

    Proteomic databases

    PaxDbiP22106.
    PRIDEiP22106.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC73768; AAC73768; b0674.
    BAA35317; BAA35317; BAA35317.
    GeneIDi945281.
    KEGGiecj:Y75_p0653.
    eco:b0674.
    PATRICi32116529. VBIEscCol129921_0697.

    Organism-specific databases

    EchoBASEiEB0090.
    EcoGeneiEG10092. asnB.

    Phylogenomic databases

    eggNOGiCOG0367.
    HOGENOMiHOG000027493.
    InParanoidiP22106.
    KOiK01953.
    OMAiDWSGIYS.
    OrthoDBiEOG6J48H7.
    PhylomeDBiP22106.

    Enzyme and pathway databases

    UniPathwayiUPA00134; UER00195.
    BioCyciEcoCyc:ASNSYNB-MONOMER.
    ECOL316407:JW0660-MONOMER.
    MetaCyc:ASNSYNB-MONOMER.
    BRENDAi6.3.5.4. 2026.

    Miscellaneous databases

    EvolutionaryTraceiP22106.
    PROiP22106.

    Gene expression databases

    GenevestigatoriP22106.

    Family and domain databases

    Gene3Di3.40.50.620. 1 hit.
    3.60.20.10. 1 hit.
    InterProiIPR006426. Asn_synth_AEB.
    IPR001962. Asn_synthase.
    IPR017932. GATase_2_dom.
    IPR000583. GATase_dom.
    IPR029055. Ntn_hydrolases_N.
    IPR014729. Rossmann-like_a/b/a_fold.
    [Graphical view]
    PfamiPF00733. Asn_synthase. 1 hit.
    PF13537. GATase_7. 1 hit.
    [Graphical view]
    SUPFAMiSSF56235. SSF56235. 1 hit.
    TIGRFAMsiTIGR01536. asn_synth_AEB. 1 hit.
    PROSITEiPS51278. GATASE_TYPE_2. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Nucleotide sequence of Escherichia coli asnB and deduced amino acid sequence of asparagine synthetase B."
      Scofield M.A., Lewis W.S., Schuster S.M.
      J. Biol. Chem. 265:12895-12902(1990) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: K12.
    2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    3. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    4. "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
      Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
      Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    5. "Glutamine-dependent nitrogen transfer in Escherichia coli asparagine synthetase B. Searching for the catalytic triad."
      Boehlein S.K., Richards N.G., Schuster S.M.
      J. Biol. Chem. 269:7450-7457(1994) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF N-TERMINUS, CLEAVAGE OF INITIATOR METHIONINE, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF CYS-2; HIS-30; ASP-34; HIS-81 AND ALA-105.
      Strain: K12.
    6. "Genetic and biomedical studies demonstrating a second gene coding for asparagine synthetase in Escherichia coli."
      Humbert R., Simoni R.D.
      J. Bacteriol. 142:212-220(1980) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION IN ASPARAGINE BIOSYNTHESIS.
      Strain: K12.
    7. "Mapping the aspartic acid binding site of Escherichia coli asparagine synthetase B using substrate analogs."
      Parr I.B., Boehlein S.K., Dribben A.B., Schuster S.M., Richards N.G.
      J. Med. Chem. 39:2367-2378(1996) [PubMed] [Europe PMC] [Abstract]
      Cited for: ENZYME REGULATION, INHIBITION STUDIES.
    8. "Revisiting the steady state kinetic mechanism of glutamine-dependent asparagine synthetase from Escherichia coli."
      Tesson A.R., Soper T.S., Ciustea M., Richards N.G.
      Arch. Biochem. Biophys. 413:23-31(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: KINETIC PARAMETERS.
      Strain: K12.
    9. "A conserved glutamate controls the commitment to acyl-adenylate formation in asparagine synthetase."
      Meyer M.E., Gutierrez J.A., Raushel F.M., Richards N.G.
      Biochemistry 49:9391-9401(2010) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF GLU-349, KINETIC MODEL.
      Strain: K12.
    10. "Three-dimensional structure of Escherichia coli asparagine synthetase B: a short journey from substrate to product."
      Larsen T.M., Boehlein S.K., Schuster S.M., Richards N.G.J., Thoden J.B., Holden H.M., Rayment I.
      Biochemistry 38:16146-16157(1999) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 2-554 OF MUTANT ALA-2 IN COMPLEX WITH AMP AND L-GLUTAMINE, SUBUNIT.
      Strain: K12.

    Entry informationi

    Entry nameiASNB_ECOLI
    AccessioniPrimary (citable) accession number: P22106
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: August 1, 1991
    Last sequence update: January 23, 2007
    Last modified: May 27, 2015
    This is version 135 of the entry and version 3 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Direct protein sequencing, Reference proteome

    Documents

    1. Escherichia coli
      Escherichia coli (strain K12): entries and cross-references to EcoGene
    2. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    3. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    4. 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.