Reviewed, UniProtKB/Swiss-Prot P00805 (ASPG2_ECOLI)

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

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.

General annotation (Comments)

Catalytic activity	L-asparagine + H2O = L-aspartate + NH3.
Subunit structure	Homotetramer. Ref.8
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: KM=0.115 µM for L-asparagine

Ontologies

Keywords
Cellular component	Periplasm
Domain	Signal
Molecular function	Hydrolase
PTM	Disulfide bond
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-SubCell
Molecular function	asparaginase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 22

22

Ref.5

Chain

23 – 348

326

L-asparaginase 2

PRO_0000002356

Sites

Active site

34

1

Ref.7 Ref.9

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

Ref.12

Experimental info

Mutagenesis

34

1

T → A: Almost no activity. Ref.10 Ref.11

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

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348

Helix Strand Turn

Details...

Beta strand

25 – 33

9

Turn

58 – 60

3

Helix

62 – 66

5

Beta strand

69 – 74

6

Helix

80 – 82

3

Helix

85 – 98

14

Helix

99 – 101

3

Beta strand

103 – 108

6

Beta strand

111 – 113

3

Helix

114 – 124

11

Beta strand

131 – 134

4

Helix

147 – 159

13

Helix

161 – 163

3

Beta strand

168 – 172

5

Beta strand

175 – 178

4

Turn

179 – 181

3

Beta strand

182 – 184

3

Beta strand

186 – 188

3

Beta strand

193 – 195

3

Turn

196 – 198

3

Beta strand

201 – 205

5

Beta strand

208 – 211

4

Helix

220 – 222

3

Beta strand

236 – 240

5

Helix

248 – 255

8

Beta strand

259 – 266

8

Turn

267 – 269

3

Helix

273 – 284

12

Beta strand

288 – 298

11

Beta strand

304 – 306

3

Helix

308 – 311

4

Helix

321 – 331

11

Turn

332 – 334

3

Helix

338 – 347

10

Sequences

	Sequence		Length	Mass (Da)
	P00805-1 [UniParc]. Last modified November 1, 1990. Version 2. Checksum: 2076987C0E8B2F99	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

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.

Web resources

Worthington enzyme manual

Cross-references

Sequence databases

EMBLGenBankDDBJ

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

PDBPDBe

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

Search...

Protein-protein interaction databases

DIP

DIP:9110N.

STRING

P00805.

Genome annotation databases

GeneID

947454.

GenomeReviews

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

KEGG

ecj:JW2924.
eco:b2957.

Organism-specific databases

EchoBASE

EB0044.

EcoGene

EG10046. ansB.

CMR

Search...

Phylogenomic databases

HOGENOM

P00805.

OMA

NFGPLGY.

Enzyme and pathway databases

BioCyc

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

Gene expression databases

Genevestigator

P00805.

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...

Entry information

Entry name

ASPG2_ECOLI

Accession

Primary (citable) accession number: P00805
Secondary accession number(s): Q2M9N6

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 21, 1986
Last sequence update:	November 1, 1990
Last modified:	November 3, 2009
This is version 107 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)