P00805 (ASPG2_ECOLI) Reviewed, UniProtKB/Swiss-Prot

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) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

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. Ref.13
Subunit structure	Homotetramer. Ref.8 Ref.9 Ref.12 Ref.13 Ref.14
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 Reference proteome
Gene Ontology (GO)
Biological_process	asparagine metabolic process Inferred from electronic annotation. Source: InterPro
Cellular_component	cytosol Inferred from direct assay PubMed 16858726. Source: UniProtKB outer membrane-bounded periplasmic space Inferred from direct assay PubMed 4962587. Source: EcoCyc
Molecular_function	asparaginase activity Inferred from direct assay PubMed 1821783. Source: EcoCyc
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

Regions

Region

80 – 81

2

Substrate binding

Region

111 – 112

2

Substrate binding

Sites

Active site

34

1

O-isoaspartyl threonine intermediate Ref.7 Ref.9 Ref.13

Amino acid modifications

Disulfide bond

99 ↔ 127

Ref.12 Ref.13

Experimental info

Mutagenesis

34

1

T → A: Loss of enzyme activity. Ref.9 Ref.10 Ref.11

Mutagenesis

111

1

T → S: Reduced enzyme activity. Ref.10 Ref.11 Ref.13

Mutagenesis

111

1

T → V: Loss of enzyme activity. Ref.10 Ref.11 Ref.13

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

Helix

34 – 36

3

Beta strand

42 – 44

3

Beta strand

48 – 50

3

Helix

54 – 59

6

Helix

62 – 66

5

Beta strand

69 – 78

10

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

302 – 306

5

Helix

308 – 311

4

Beta strand

313 – 315

3

Helix

321 – 331

11

Turn

332 – 334

3

Helix

338 – 345

8

Sequences

	Sequence		Length	Mass (Da)
	P00805 [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] [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]	"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] [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]	"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.
+	Additional computationally mapped references.

Web resources

Worthington enzyme manual

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

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

NP_417432.1. NC_000913.2.
YP_491156.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

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	[»]

ProteinModelPortal

P00805.

SMR

P00805. Positions 23-348.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-9110N.

IntAct

P00805. 4 interactions.

STRING

511145.b2957.

Protein family/group databases

Allergome

8365. Esc c Asparaginase.

Proteomic databases

PaxDb

P00805.

PRIDE

P00805.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC75994; AAC75994; b2957.
BAE77020; BAE77020; BAE77020.

GeneID

12930249.
947454.

KEGG

ecj:Y75_p2887.
eco:b2957.

PATRIC

32121326. VBIEscCol129921_3051.

Organism-specific databases

EchoBASE

EB0044.

EcoGene

EG10046. ansB.

Phylogenomic databases

eggNOG

COG0252.

HOGENOM

HOG000044165.

KO

K01424.

OMA

GEIDDKA.

ProtClustDB

PRK11096.

Enzyme and pathway databases

BioCyc

EcoCyc:ANSB-MONOMER.
ECOL316407:JW2924-MONOMER.
MetaCyc:ANSB-MONOMER.

SABIO-RK

P00805.

Gene expression databases

Genevestigator

P00805.

Family and domain databases

Gene3D

3.40.50.1170. 1 hit.
3.40.50.40. 1 hit.

InterPro

IPR004550. AsnASE_II.
IPR006034. Asparaginase/glutaminase.
IPR020827. Asparaginase/glutaminase_AS1.
IPR027475. Asparaginase/glutaminase_AS2.
IPR027473. L-asparaginase_C.
IPR027474. L-asparaginase_N.
[Graphical view]

Pfam

PF00710. Asparaginase. 1 hit.
[Graphical view]

PIRSF

PIRSF001220. L-ASNase_gatD. 1 hit.

PRINTS

PR00139. ASNGLNASE.

SMART

SM00870. Asparaginase. 1 hit.
[Graphical view]

SUPFAM

SSF53774. Asp/Glutamnse. 1 hit.

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...

Other

EvolutionaryTrace

P00805.

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:	May 29, 2013
This is version 140 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program