Skip Header

You are using a version of Internet Explorer that may not display all features of this website. Please upgrade to a modern browser.
Contribute Send feedback
Read comments (?) or add your own

P22106 (ASNB_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified July 9, 2014. Version 129. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (4) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Interactions·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Asparagine synthetase B [glutamine-hydrolyzing]

Short name=AS-B
EC=6.3.5.4
Gene names
Name:asnB
Ordered Locus Names:b0674, JW0660
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length554 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

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. Ref.5 Ref.6 Ref.9

Catalytic activity

ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate. Ref.5 Ref.9

Enzyme regulation

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. Ref.7

Pathway

Amino-acid biosynthesis; L-asparagine biosynthesis; L-asparagine from L-aspartate (L-Gln route): step 1/1.

Subunit structure

Homodimer. Ref.10

Sequence similarities

Belongs to the asparagine synthetase family.

Contains 1 glutamine amidotransferase type-2 domain.

Biophysicochemical properties

Kinetic parameters:

KM=17 mM for ammonia Ref.5 Ref.8 Ref.9

KM=0.53 mM for aspartate (when assaying the ammonia-dependent synthetase reaction)

KM=0.85 mM for aspartate (when assaying the glutamine-dependent synthetase reaction)

KM=0.26 mM for ATP (when assaying the glutamine-dependent synthetase reaction)

KM=0.66 mM for glutamine (when assaying the glutamine-dependent synthetase reaction)

pH dependence:

Optimum pH is 6.5-8.

Binary interactions

With

Entry

#Exp.

IntAct

Notes

itself3EBI-549123,EBI-549123

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Initiator methionine11Removed Ref.5
Chain2 – 554553Asparagine synthetase B [glutamine-hydrolyzing]
PRO_0000056931

Regions

Domain2 – 186185Glutamine amidotransferase type-2
Nucleotide binding347 – 3482ATP
Region50 – 545Glutamine binding
Region75 – 773Glutamine binding

Sites

Active site21For GATase activity
Binding site991Glutamine
Binding site2331ATP; via carbonyl oxygen
Binding site2731ATP; via amide nitrogen and carbonyl oxygen
Site3491Important for beta-aspartyl-AMP intermediate formation

Experimental info

Mutagenesis21C → A or S: Loss of glutamine-dependent activity but no effect on ammonia-dependent asparagine synthetase activity. Ref.5
Mutagenesis301H → A: 4,5-fold decrease in glutamine affinity. Ref.5
Mutagenesis341D → N or E: Little effect on the kinetic properties. Ref.5
Mutagenesis811H → A: 5-fold decrease in glutamine affinity. Ref.5
Mutagenesis1051A → H: Little effect on the kinetic properties. Ref.5
Mutagenesis3491E → A or Q: Loss of glutamine- and ammonia-dependent synthetase activity, but still exhibits glutaminase activity. Ref.9
Mutagenesis3491E → 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. Ref.9

Secondary structure

................................................................................................ 554
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P22106 [UniParc].

Last modified January 23, 2007. Version 3.
Checksum: 9091B269112C9F5C

FASTA55462,659
        10         20         30         40         50         60 
MCSIFGVFDI KTDAVELRKK ALELSRLMRH RGPDWSGIYA SDNAILAHER LSIVDVNAGA 

        70         80         90        100        110        120 
QPLYNQQKTH VLAVNGEIYN HQALRAEYGD RYQFQTGSDC EVILALYQEK GPEFLDDLQG 

       130        140        150        160        170        180 
MFAFALYDSE KDAYLIGRDH LGIIPLYMGY DEHGQLYVAS EMKALVPVCR TIKEFPAGSY 

       190        200        210        220        230        240 
LWSQDGEIRS YYHRDWFDYD AVKDNVTDKN ELRQALEDSV KSHLMSDVPY GVLLSGGLDS 

       250        260        270        280        290        300 
SIISAITKKY AARRVEDQER SEAWWPQLHS FAVGLPGSPD LKAAQEVANH LGTVHHEIHF 

       310        320        330        340        350        360 
TVQEGLDAIR DVIYHIETYD VTTIRASTPM YLMSRKIKAM GIKMVLSGEG SDEVFGGYLY 

       370        380        390        400        410        420 
FHKAPNAKEL HEETVRKLLA LHMYDCARAN KAMSAWGVEA RVPFLDKKFL DVAMRINPQD 

       430        440        450        460        470        480 
KMCGNGKMEK HILRECFEAY LPASVAWRQK EQFSDGVGYS WIDTLKEVAA QQVSDQQLET 

       490        500        510        520        530        540 
ARFRFPYNTP TSKEAYLYRE IFEELFPLPS AAECVPGGPS VACSSAKAIE WDEAFKKMDD 

       550 
PSGRAVGVHQ SAYK 

« Hide

References

« Hide 'large scale' references
[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]"A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12.7-28.0 min region on the linkage map."
Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S., Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K. expand/collapse author list , Mori H., Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G., Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M., Horiuchi T.
DNA Res. 3:137-155(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[3]"The complete genome sequence of Escherichia coli K-12."
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.
Science 277:1453-1462(1997) [PubMed] [Europe PMC] [Abstract]
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.
[11]Erratum
Larsen T.M., Boehlein S.K., Schuster S.M., Richards N.G.J., Thoden J.B., Holden H.M., Rayment I.
Biochemistry 39:7330-7330(2000) [PubMed] [Europe PMC] [Abstract]
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
J05554 Genomic DNA. Translation: AAA23498.1.
U00096 Genomic DNA. Translation: AAC73768.1.
AP009048 Genomic DNA. Translation: BAA35317.1.
PIRAJECN. A36616.
RefSeqNP_415200.1. NC_000913.3.
YP_488954.1. NC_007779.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1CT9X-ray2.00A/B/C/D2-554[»]
ProteinModelPortalP22106.
SMRP22106. Positions 3-517.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

DIPDIP-9177N.
IntActP22106. 7 interactions.
MINTMINT-1306603.
STRING511145.b0674.

Chemistry

DrugBankDB00131. Adenosine monophosphate.

Protein family/group databases

MEROPSC44.976.

Proteomic databases

PaxDbP22106.
PRIDEP22106.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC73768; AAC73768; b0674.
BAA35317; BAA35317; BAA35317.
GeneID12932818.
945281.
KEGGecj:Y75_p0653.
eco:b0674.
PATRIC32116529. VBIEscCol129921_0697.

Organism-specific databases

EchoBASEEB0090.
EcoGeneEG10092. asnB.

Phylogenomic databases

eggNOGCOG0367.
HOGENOMHOG000027493.
KOK01953.
OMAWSGIYSS.
OrthoDBEOG6J48H7.
PhylomeDBP22106.

Enzyme and pathway databases

BioCycEcoCyc:ASNSYNB-MONOMER.
ECOL316407:JW0660-MONOMER.
MetaCyc:ASNSYNB-MONOMER.
UniPathwayUPA00134; UER00195.

Gene expression databases

GenevestigatorP22106.

Family and domain databases

Gene3D3.40.50.620. 1 hit.
3.60.20.10. 1 hit.
InterProIPR006426. 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]
PfamPF00733. Asn_synthase. 1 hit.
PF13537. GATase_7. 1 hit.
[Graphical view]
PIRSFPIRSF001589. Asn_synthetase_glu-h. 1 hit.
SUPFAMSSF56235. SSF56235. 1 hit.
TIGRFAMsTIGR01536. asn_synth_AEB. 1 hit.
PROSITEPS51278. GATASE_TYPE_2. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP22106.
PROP22106.

Entry information

Entry nameASNB_ECOLI
AccessionPrimary (citable) accession number: P22106
Entry history
Integrated into UniProtKB/Swiss-Prot: August 1, 1991
Last sequence update: January 23, 2007
Last modified: July 9, 2014
This is version 129 of the entry and version 3 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene