L-asparaginase 2 precursor - Escherichia coli (strain K12)

Names and origin

Protein names

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 names

Name:	ansB
Ordered Locus Names:	b2957, JW2924

Organism

Escherichia coli (strain K12) [Complete proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

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Protein attributes

Sequence length	348 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level.

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General annotation (Comments)

Catalytic activity	L-asparagine + H(2)O = L-aspartate + NH(3).
Subunit structure	Homotetramer.
Subcellular location	Periplasm.
Induction	By cAMP and anaerobiosis.
Pharmaceutical use	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.
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.
Sequence similarities	Belongs to the asparaginase 1 family.
Biophysicochemical properties	Kinetic parameters: K_M=0.115 µM for L-asparagine

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Ontologies

Keywords
Cellular component	Periplasm
Domain	Signal
Molecular function	Hydrolase
Technical term	3D-structure Complete proteome Direct protein sequencing Pharmaceutical
Gene Ontology (GO)
Biological process	asparagine metabolic process Inferred from electronic annotation. Source: InterPro
Cellular component	cytosol Inferred from direct assay. Source: UniProtKB periplasmic space Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	asparaginase activity Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

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Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 22

22

Chain

23 – 348

326

L-asparaginase 2

PRO_0000002356

Sites

Active site

34

1

Active site

111

1

By similarity

Active site

112

1

By similarity

Active site

184

1

By similarity

Binding site

141

1

Substrate

Amino acid modifications

Disulfide bond

99 ↔ 127

Experimental info

Mutagenesis

34

1

T → A: Almost no activity

Sequence conflict

49

1

V → A AA sequence Ref.5

Sequence conflict

86

1

N → D AA sequence Ref.5

Sequence conflict

132

1

Missing AA sequence Ref.5

Sequence conflict

156

1

Missing AA sequence Ref.5

Sequence conflict

171

1

Missing AA sequence Ref.5

Sequence conflict

206

1

N → D AA sequence Ref.5

Sequence conflict

268

1

N → D AA sequence Ref.5

Sequence conflict

274

1

S → T AA sequence Ref.5

Sequence conflict

285

1

T → D AA sequence Ref.5

Sequence conflict

290

1

Missing AA sequence Ref.5

Sequence conflict

330

1

Missing AA sequence Ref.5

Secondary structure

1

.

348

Helix Strand Turn

Details...

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Sequences

Sequence

Length

Mass (Da)

Tools

P00805-1 [UniParc].

Last modified November 1, 1990. Version 2.
Checksum: 2076987C0E8B2F99
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FASTA

348

36,851

        10         20         30         40         50         60 
MEFFKKTALA ALVMGFSGAA LALPNITILA TGGTIAGGGD SATKSNYTVG KVGVENLVNA 

        70         80         90        100        110        120 
VPQLKDIANV KGEQVVNIGS QDMNDNVWLT LAKKINTDCD KTDGFVITHG TDTMEETAYF 

       130        140        150        160        170        180 
LDLTVKCDKP VVMVGAMRPS TSMSADGPFN LYNAVVTAAD KASANRGVLV VMNDTVLDGR 

       190        200        210        220        230        240 
DVTKTNTTDV ATFKSVNYGP LGYIHNGKID YQRTPARKHT SDTPFDVSKL NELPKVGIVY 

       250        260        270        280        290        300 
NYANASDLPA KALVDAGYDG IVSAGVGNGN LYKSVFDTLA TAAKTGTAVV RSSRVPTGAT 

       310        320        330        340 
TQDAEVDDAK YGFVASGTLN PQKARVLLQL ALTQTKDPQQ IQQIFNQY

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References

	« Hide 'large scale' referencesShow 'large scale' references »
[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: 2407723] [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: 2144836] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[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 277:1453-1474(1997) [PubMed: 9278503] [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: 16738553] [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: 6766894] [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: 387570] [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: 321449] [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: 4561256] [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: 1906013] [Abstract] Cited for: ACTIVE SITE THR-34.
[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: 1521538] [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: 1287659] [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: 8434007] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
[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: 8706862] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT VAL-111.
+	Additional computationally mapped references.

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Web resources

Worthington enzyme manual

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Cross-references

Sequence databases

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.

PIR

XDEC. A35132.

RefSeq

AP_003514.1.
NP_417432.1.

3D structure databases

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1HO3	X-ray	2.50	A/B	23-348	[»]
1IHD	X-ray	2.65	A/C	23-348	[»]
1JAZ	X-ray	2.27	A/B	23-348	[»]
1JJA	X-ray	2.30	A/B/C/D/E/F	23-348	[»]
1NNS	X-ray	1.95	A/B	23-348	[»]
3ECA	X-ray	2.40	A/B/C/D	23-348	[»]
4ECA	X-ray	2.20	A/B/C/D	23-348	[»]

ModBase

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Protein-protein interaction databases

DIP

DIP:9110N.

Genome annotation databases

GeneID

947454.

GenomeReviews

Gene locus b2957 in contig U00096_GR.
Gene locus JW2924 in contig AP009048_GR.

KEGG

ecj:JW2924.
eco:b2957.

Organism-specific databases

EchoBASE

EB0044.

EcoGene

EG10046. ansB.

CMR

Search...

Phylogenomic databases

HOGENOM

P00805.

Enzyme and pathway databases

BioCyc

EcoCyc:ANSB-MON.
MetaCyc:ANSB-MON.

Family and domain databases

InterPro

IPR004550. AsnASE_II.
IPR006034. Asp/Glutamnse.
[Graphical view]

PANTHER

PTHR11707. Asp/Glutamnse. 1 hit.

Pfam

PF00710. Asparaginase. 1 hit.
[Graphical view]

PRINTS

PR00139. ASNGLNASE.

ProDom

PD003221. Asp/Glutamnse. 1 hit.
[Graphical view] [Entries sharing at least one domain]

TIGRFAMs

TIGR00520. asnASE_II. 1 hit.

PROSITE

PS00144. ASN_GLN_ASE_1. 1 hit.
PS00917. ASN_GLN_ASE_2. 1 hit.
[Graphical view]

ProtoNet

Search...

Names and origin

Protein attributes

General annotation (Comments)

Ontologies

Keywords

Gene Ontology (GO)

Sequence annotation (Features)

Molecule processing

Sites

Amino acid modifications

Experimental info

Secondary structure

Sequences

References

Web resources

Cross-references

Sequence databases

3D structure databases

Protein-protein interaction databases

Genome annotation databases

Organism-specific databases

Phylogenomic databases

Enzyme and pathway databases

Family and domain databases

Other Resources