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

L-asparaginase 2

Gene

ansB

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

Functioni

Catalytic activityi

L-asparagine + H2O = L-aspartate + NH3.1 Publication

Kineticsi

  1. KM=0.115 µM for L-asparagine

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Active sitei34 – 341O-isoaspartyl threonine intermediatePROSITE-ProRule annotation1 Publication

    GO - Molecular functioni

    • asparaginase activity Source: EcoCyc
    • identical protein binding Source: IntAct

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Hydrolase

    Enzyme and pathway databases

    BioCyciEcoCyc:ANSB-MONOMER.
    ECOL316407:JW2924-MONOMER.
    MetaCyc:ANSB-MONOMER.
    BRENDAi3.5.1.1. 2026.
    SABIO-RKP00805.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    L-asparaginase 2 (EC:3.5.1.1)
    Alternative name(s):
    L-asparaginase II
    Short name:
    L-ASNase II
    L-asparagine amidohydrolase II
    INN: Colaspase
    Gene namesi
    Name:ansB
    Ordered Locus Names:b2957, JW2924
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG10046. ansB.

    Subcellular locationi

    GO - Cellular componenti

    • cytosol Source: UniProtKB
    • outer membrane-bounded periplasmic space Source: EcoCyc
    • periplasmic space Source: EcoliWiki
    Complete GO annotation...

    Keywords - Cellular componenti

    Periplasm

    Pathology & Biotechi

    Pharmaceutical usei

    Available under the names Crastinin (Bayer), Elspar (Merck), Kidrolase (Rhone-Poulenc) and Leunase (Kyowa). Also available as a PEG-conjugated form (Pegaspargase) under the name Oncaspar (Enzon). Used as an antineoplastic in chemotherapy. Reduces the quantity of asparagine available to cancer cells.

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi34 – 341T → A: Loss of enzyme activity. 1 Publication
    Mutagenesisi111 – 1111T → S: Reduced enzyme activity. 1 Publication
    Mutagenesisi111 – 1111T → V: Loss of enzyme activity. 1 Publication

    Protein family/group databases

    Allergomei8365. Esc c Asparaginase.

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Signal peptidei1 – 22221 PublicationAdd
    BLAST
    Chaini23 – 348326L-asparaginase 2PRO_0000002356Add
    BLAST

    Amino acid modifications

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Disulfide bondi99 ↔ 1272 Publications

    Keywords - PTMi

    Disulfide bond

    Proteomic databases

    PaxDbiP00805.
    PRIDEiP00805.

    Expressioni

    Inductioni

    By cAMP and anaerobiosis.

    Gene expression databases

    GenevestigatoriP00805.

    Interactioni

    Subunit structurei

    Homotetramer.5 Publications

    Binary interactionsi

    WithEntry#Exp.IntActNotes
    itself2EBI-907458,EBI-907458

    Protein-protein interaction databases

    DIPiDIP-9110N.
    IntActiP00805. 7 interactions.
    STRINGi511145.b2957.

    Structurei

    Secondary structure

    1
    348
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi25 – 339Combined sources
    Helixi34 – 363Combined sources
    Beta strandi42 – 443Combined sources
    Beta strandi48 – 503Combined sources
    Helixi54 – 596Combined sources
    Helixi62 – 665Combined sources
    Beta strandi69 – 7810Combined sources
    Helixi80 – 823Combined sources
    Helixi85 – 9814Combined sources
    Helixi99 – 1013Combined sources
    Beta strandi103 – 1086Combined sources
    Beta strandi111 – 1133Combined sources
    Helixi114 – 12411Combined sources
    Beta strandi131 – 1344Combined sources
    Helixi147 – 15913Combined sources
    Helixi161 – 1633Combined sources
    Beta strandi168 – 1725Combined sources
    Beta strandi175 – 1784Combined sources
    Turni179 – 1813Combined sources
    Beta strandi182 – 1843Combined sources
    Beta strandi186 – 1883Combined sources
    Beta strandi193 – 1953Combined sources
    Turni196 – 1983Combined sources
    Beta strandi201 – 2055Combined sources
    Beta strandi208 – 2114Combined sources
    Helixi220 – 2223Combined sources
    Beta strandi236 – 2405Combined sources
    Helixi248 – 2558Combined sources
    Beta strandi259 – 2668Combined sources
    Turni267 – 2693Combined sources
    Helixi273 – 28412Combined sources
    Beta strandi288 – 29811Combined sources
    Beta strandi302 – 3065Combined sources
    Helixi308 – 3114Combined sources
    Beta strandi313 – 3153Combined sources
    Helixi321 – 33111Combined sources
    Turni332 – 3343Combined sources
    Helixi338 – 3458Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1HO3X-ray2.50A/B23-348[»]
    1IHDX-ray2.65A/C23-348[»]
    1JAZX-ray2.27A/B23-348[»]
    1JJAX-ray2.30A/B/C/D/E/F23-348[»]
    1NNSX-ray1.95A/B23-348[»]
    3ECAX-ray2.40A/B/C/D23-348[»]
    4ECAX-ray2.20A/B/C/D23-348[»]
    ProteinModelPortaliP00805.
    SMRiP00805. Positions 23-348.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP00805.

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini24 – 348325Asparaginase/glutaminasePROSITE-ProRule annotationAdd
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni80 – 812Substrate binding
    Regioni111 – 1122Substrate binding

    Sequence similaritiesi

    Belongs to the asparaginase 1 family.Curated
    Contains 1 asparaginase/glutaminase domain.PROSITE-ProRule annotation

    Keywords - Domaini

    Signal

    Phylogenomic databases

    eggNOGiCOG0252.
    HOGENOMiHOG000044165.
    InParanoidiP00805.
    KOiK01424.
    OMAiFQSPNFG.
    OrthoDBiEOG6VTK47.
    PhylomeDBiP00805.

    Family and domain databases

    Gene3Di3.40.50.1170. 1 hit.
    3.40.50.40. 1 hit.
    InterProiIPR004550. AsnASE_II.
    IPR006034. Asparaginase/glutaminase.
    IPR020827. Asparaginase/glutaminase_AS1.
    IPR027475. Asparaginase/glutaminase_AS2.
    IPR027473. L-asparaginase_C.
    IPR027474. L-asparaginase_N.
    [Graphical view]
    PfamiPF00710. Asparaginase. 1 hit.
    [Graphical view]
    PIRSFiPIRSF001220. L-ASNase_gatD. 1 hit.
    PRINTSiPR00139. ASNGLNASE.
    SMARTiSM00870. Asparaginase. 1 hit.
    [Graphical view]
    SUPFAMiSSF53774. SSF53774. 1 hit.
    TIGRFAMsiTIGR00520. asnASE_II. 1 hit.
    PROSITEiPS00144. ASN_GLN_ASE_1. 1 hit.
    PS00917. ASN_GLN_ASE_2. 1 hit.
    PS51732. ASN_GLN_ASE_3. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

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

    P00805-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MEFFKKTALA ALVMGFSGAA LALPNITILA TGGTIAGGGD SATKSNYTVG
    60 70 80 90 100
    KVGVENLVNA VPQLKDIANV KGEQVVNIGS QDMNDNVWLT LAKKINTDCD
    110 120 130 140 150
    KTDGFVITHG TDTMEETAYF LDLTVKCDKP VVMVGAMRPS TSMSADGPFN
    160 170 180 190 200
    LYNAVVTAAD KASANRGVLV VMNDTVLDGR DVTKTNTTDV ATFKSVNYGP
    210 220 230 240 250
    LGYIHNGKID YQRTPARKHT SDTPFDVSKL NELPKVGIVY NYANASDLPA
    260 270 280 290 300
    KALVDAGYDG IVSAGVGNGN LYKSVFDTLA TAAKTGTAVV RSSRVPTGAT
    310 320 330 340
    TQDAEVDDAK YGFVASGTLN PQKARVLLQL ALTQTKDPQQ IQQIFNQY
    Length:348
    Mass (Da):36,851
    Last modified:November 1, 1990 - v2
    Checksum:i2076987C0E8B2F99
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti49 – 491V → A AA sequence (PubMed:6766894).Curated
    Sequence conflicti86 – 861N → D AA sequence (PubMed:6766894).Curated
    Sequence conflicti132 – 1321Missing AA sequence (PubMed:6766894).Curated
    Sequence conflicti156 – 1561Missing AA sequence (PubMed:6766894).Curated
    Sequence conflicti171 – 1711Missing AA sequence (PubMed:6766894).Curated
    Sequence conflicti206 – 2061N → D AA sequence (PubMed:6766894).Curated
    Sequence conflicti268 – 2681N → D AA sequence (PubMed:6766894).Curated
    Sequence conflicti274 – 2741S → T AA sequence (PubMed:6766894).Curated
    Sequence conflicti285 – 2851T → D AA sequence (PubMed:6766894).Curated
    Sequence conflicti290 – 2901Missing AA sequence (PubMed:6766894).Curated
    Sequence conflicti330 – 3301Missing AA sequence (PubMed:6766894).Curated

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    M34277 Genomic DNA. Translation: AAA24062.1.
    M34234 Genomic DNA. Translation: AAA23445.1.
    U28377 Genomic DNA. Translation: AAA69124.1.
    U00096 Genomic DNA. Translation: AAC75994.1.
    AP009048 Genomic DNA. Translation: BAE77020.1.
    PIRiA35132. XDEC.
    RefSeqiNP_417432.1. NC_000913.3.

    Genome annotation databases

    EnsemblBacteriaiAAC75994; AAC75994; b2957.
    BAE77020; BAE77020; BAE77020.
    GeneIDi947454.
    KEGGiecj:Y75_p2887.
    eco:b2957.
    PATRICi32121326. VBIEscCol129921_3051.

    Cross-referencesi

    Web resourcesi

    Worthington enzyme manual

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    M34277 Genomic DNA. Translation: AAA24062.1.
    M34234 Genomic DNA. Translation: AAA23445.1.
    U28377 Genomic DNA. Translation: AAA69124.1.
    U00096 Genomic DNA. Translation: AAC75994.1.
    AP009048 Genomic DNA. Translation: BAE77020.1.
    PIRiA35132. XDEC.
    RefSeqiNP_417432.1. NC_000913.3.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1HO3X-ray2.50A/B23-348[»]
    1IHDX-ray2.65A/C23-348[»]
    1JAZX-ray2.27A/B23-348[»]
    1JJAX-ray2.30A/B/C/D/E/F23-348[»]
    1NNSX-ray1.95A/B23-348[»]
    3ECAX-ray2.40A/B/C/D23-348[»]
    4ECAX-ray2.20A/B/C/D23-348[»]
    ProteinModelPortaliP00805.
    SMRiP00805. Positions 23-348.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-9110N.
    IntActiP00805. 7 interactions.
    STRINGi511145.b2957.

    Protein family/group databases

    Allergomei8365. Esc c Asparaginase.

    Proteomic databases

    PaxDbiP00805.
    PRIDEiP00805.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC75994; AAC75994; b2957.
    BAE77020; BAE77020; BAE77020.
    GeneIDi947454.
    KEGGiecj:Y75_p2887.
    eco:b2957.
    PATRICi32121326. VBIEscCol129921_3051.

    Organism-specific databases

    EchoBASEiEB0044.
    EcoGeneiEG10046. ansB.

    Phylogenomic databases

    eggNOGiCOG0252.
    HOGENOMiHOG000044165.
    InParanoidiP00805.
    KOiK01424.
    OMAiFQSPNFG.
    OrthoDBiEOG6VTK47.
    PhylomeDBiP00805.

    Enzyme and pathway databases

    BioCyciEcoCyc:ANSB-MONOMER.
    ECOL316407:JW2924-MONOMER.
    MetaCyc:ANSB-MONOMER.
    BRENDAi3.5.1.1. 2026.
    SABIO-RKP00805.

    Miscellaneous databases

    EvolutionaryTraceiP00805.
    PROiP00805.

    Gene expression databases

    GenevestigatoriP00805.

    Family and domain databases

    Gene3Di3.40.50.1170. 1 hit.
    3.40.50.40. 1 hit.
    InterProiIPR004550. AsnASE_II.
    IPR006034. Asparaginase/glutaminase.
    IPR020827. Asparaginase/glutaminase_AS1.
    IPR027475. Asparaginase/glutaminase_AS2.
    IPR027473. L-asparaginase_C.
    IPR027474. L-asparaginase_N.
    [Graphical view]
    PfamiPF00710. Asparaginase. 1 hit.
    [Graphical view]
    PIRSFiPIRSF001220. L-ASNase_gatD. 1 hit.
    PRINTSiPR00139. ASNGLNASE.
    SMARTiSM00870. Asparaginase. 1 hit.
    [Graphical view]
    SUPFAMiSSF53774. SSF53774. 1 hit.
    TIGRFAMsiTIGR00520. asnASE_II. 1 hit.
    PROSITEiPS00144. ASN_GLN_ASE_1. 1 hit.
    PS00917. ASN_GLN_ASE_2. 1 hit.
    PS51732. ASN_GLN_ASE_3. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Analysis of the Escherichia coli gene encoding L-asparaginase II, ansB, and its regulation by cyclic AMP receptor and FNR proteins."
      Jennings M.P., Beacham I.R.
      J. Bacteriol. 172:1491-1498(1990) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    2. "L-asparaginase II of Escherichia coli K-12: cloning, mapping and sequencing of the ansB gene."
      Bonthron D.T.
      Gene 91:101-105(1990) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: K12.
    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. "The primary structure of L-asparaginase from Escherichia coli."
      Maita T., Matsuda G.
      Hoppe-Seyler's Z. Physiol. Chem. 361:105-117(1980) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 23-348.
    6. "Amino acid sequences of the tryptic peptides from carboxymethylated L-asparaginase from Escherichia coli."
      Maita T., Morokuma K., Matsuda G.
      Hoppe-Seyler's Z. Physiol. Chem. 360:1483-1495(1979) [PubMed] [Europe PMC] [Abstract]
      Cited for: PARTIAL PROTEIN SEQUENCE.
    7. "Structure of peptide from active site region of Escherichia coli L-asparaginase."
      Peterson R.G., Richards F.F., Handschumacher R.E.
      J. Biol. Chem. 252:2072-2076(1977) [PubMed] [Europe PMC] [Abstract]
      Cited for: ACTIVE SITE.
    8. "Chemical evidence for identical subunits in L-asparaginase from Escherichia coli B."
      Greenquist A.C., Wriston J.C. Jr.
      Arch. Biochem. Biophys. 152:280-286(1972) [PubMed] [Europe PMC] [Abstract]
      Cited for: SUBUNIT.
    9. "A catalytic role for threonine-12 of E. coli asparaginase II as established by site-directed mutagenesis."
      Harms E., Wehner A., Aung H.P., Roehm K.H.
      FEBS Lett. 285:55-58(1991) [PubMed] [Europe PMC] [Abstract]
      Cited for: ACTIVE SITE THR-34, MUTAGENESIS OF THR-34, SUBUNIT.
    10. "Site-specific mutagenesis of Escherichia coli asparaginase II. None of the three histidine residues is required for catalysis."
      Wehner A., Harms E., Jennings M.P., Beacham I.R., Derst C., Bast P., Roehm K.H.
      Eur. J. Biochem. 208:475-480(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: MUTAGENESIS OF HISTIDINE RESIDUES.
    11. "Probing the role of threonine and serine residues of E. coli asparaginase II by site-specific mutagenesis."
      Ders C., Henseling J., Roehm K.H.
      Protein Eng. 5:785-789(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: MUTAGENESIS OF THREONINE AND SERINE RESIDUES.
    12. "Crystal structure of Escherichia coli L-asparaginase, an enzyme used in cancer therapy."
      Swain A.L., Jaskolski M., Housset D., Rao J.K.M., Wlodawer A.
      Proc. Natl. Acad. Sci. U.S.A. 90:1474-1478(1993) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) IN COMPLEX WITH ASPARTIC ACID, DISULFIDE BOND, SUBUNIT.
    13. "A covalently bound catalytic intermediate in Escherichia coli asparaginase: crystal structure of a Thr-89-Val mutant."
      Palm G.J., Lubkowski J., Derst C., Schleper S., Roehm K.H., Wlodawer A.
      FEBS Lett. 390:211-216(1996) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT VAL-111, ACTIVE SITE, MUTAGENESIS OF THR-111, CATALYTIC ACTIVITY, SUBUNIT, DISULFIDE BOND.
    14. "Structural comparison of Escherichia coli L-asparaginase in two monoclinic space groups."
      Sanches M., Barbosa J.A., de Oliveira R.T., Abrahao Neto J., Polikarpov I.
      Acta Crystallogr. D 59:416-422(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF 23-348 IN COMPLEXES WITH ASPARTIC ACID, SUBUNIT.

    Entry informationi

    Entry nameiASPG2_ECOLI
    AccessioniPrimary (citable) accession number: P00805
    Secondary accession number(s): Q2M9N6
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: July 21, 1986
    Last sequence update: November 1, 1990
    Last modified: May 27, 2015
    This is version 152 of the entry and version 2 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    E.coli contains two L-asparaginase isoenzymes: L-asparaginase I, a low-affinity enzyme located in the cytoplasm, and L-asparaginase II, a high-affinity secreted enzyme.

    Keywords - Technical termi

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

    Documents

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